Frequently Asked Questions

At Tsingke, our strength lies in our fully integrated gene synthesis value chain. From intelligent design and large-scale DNA synthesis, to dual QC verification, vector construction, and downstream applications, every step is under our control. This all-in-one capability ensures accuracy, reliability, and efficiency, helping our customers speed up their scientific innovation.

The common purification methods are DSL, OPC, PAGE, HPLC and Dual PAGE & HPLC.

It will not degrade, and the dry primer can be stably stored at room temperature for at least two weeks. The average transport time is usually 5days, so the primers you receive will not degrade.

According to the purpose of the experiment. In general, about 20 bases of primer 2 OD in PCR amplification can do 400 rxns standard PCR reactions of 50 μL system. If it is to do gene splicing or annealing to do the ligation, 1 OD is enough. 

(1) Primer length generally between 15~30 bases.

(2) Primer GC content between 40% and 60%.

(3) Bases should be randomly distributed.

(4) There should be no complementary sequences in the primers themselves and between the primers

(5) The primers should have relatively high ∆G values at the 5’end and middle ∆G values, and low ∆G values at the 3’ end.

(6) The single strand of the amplification product should not form a secondary structure.

(7) The primers should have specificity.

First check the baseline-corrected or ROX-corrected raw curves. Low signal values in the amplification curves are mostly due to high background fluorescence. In probe-based detection, high background fluorescence is often caused by poor probe design, leading to insufficient quenching by the quencher group, improper pairing of the reporter and quencher groups, or low fluorescence labeling efficiency of the probe.

Oligo synthesis utilizes solid-phase phosphoramidite chemistry. The main types of oligonucleotides we synthesize include:

(1) DNA oligos;

(2) RNA oligos;

(3) Probes;

(4) siRNA;

(5) sgRNA;

(6) miRNA;

(7) ASOs;

(8) Various modified oligos.

Crude oligonucleotides contain the desired product, failure sequences and chemically modified by-products.

Oligo primers in freeze-dried state are highly stable and can be stored at -20°C for at least one year. After reconstitution, dissolved oligos can be stored as a 100 μmol/L stock solution, aliquoted into multiple tubes, and kept at -20°C for at least six months (frequent freeze-thaw cycles may reduce shelf life). Before use, dilute the concentrated solution to the working concentration for your experiments.

Each OD of primer needs to be diluted with water to a concentration of 100 μmol/L (equivalent to 100 pmol/μL).You could add an appropriate amount of nuclease-free double-distilled water (pH >6.0) or TE buffer (pH 7.5-8.0) according to your experimental needs.

Before opening the vial, centrifuge it at 3000-4000 rpm for 1 minute to prevent the loss of primers when opening.

If the pH of the water you lysed the primers in is too low or contaminated with bacteria or nucleases, it will degrade the primers. Insufficient thawing and mixing of the liquid when using may also cause inaccurate primer addition. It is recommended to split the primers, avoid repeated freeze-thawing, and use 10 mM Tris pH 7.5 buffer to dissolve the primers. There is also a possibility that the primers are not faulty, but that the quality of the materials used for PCR, especially the template, is not exactly the same as previously used.

When combining long chain primers, more reagents are required, especially primers longer than 90 base. Due to the increase in cost, the price is higher.

The entire process from synthesis, QC to packaging can be highly customized. Customizable options include, but are not limited to:

(1) Packaging in 1.5 mL, 15 mL, or 50 mL tubes

(2) Capillary electrophoresis analysis

(3) High-performance liquid chromatography (HPLC) analysis

(4) Polyacrylamide gel electrophoresis (PAGE) analysis

(5) Quantitative PCR (qPCR) analysis

(6) Full-wavelength scanning

(7) OD260/280 measurement

(8) Provision of 1 OD sample with each shipment

(9) Specialized consumables

(10) Customized Certificate of Analysis (COA)

 Synthetic primers may be yellowish brown, white or transparent, which is related to the base composition of the primers and the preparation process of synthesis, primers with high A and G content in the sequence and large OD value are usually yellowish brown, so the yellowish brown primers will not have any effect on the experiment.

A common method used in laboratories is the PAGE. For electrophoresis, use a polyacrylamide gel with 7 M urea, 20% polyacrylamide gel for primers with less than 12 bases, 16% polyacrylamide gel for primers with 12-60 bases, and 12% polyacrylamide gel for primers with more than 60 bases. Take 0.2-0.5 OD primer, dissolve with urea saturated solution or add urea dry powder into primer solution until saturated, and heat denaturation (95 ℃, 2 min) before loading sample. The purpose of adding urea is to denaturate, and to increase the specific gravity of the sample, which is easy to add the sample. After a certain period of time (about 2-3 hours), strip the polyacrylamide gel and detect the band type with a fluorescent TLC plate under the UV lamp. There is no impurity under the main band, indicating that the purity is good.

(Sometimes due to insufficient denaturation, there may be bands above the main band, which are primer secondary structure bands.)

The OD value of the synthetic primer was determined by using an ultraviolet spectrophotometer with a wavelength of 260 nm and a quartz colorimetric cup with an optical path of 1 cm to determine the optical density of the solution. The optical density of the solution should be diluted to 0.2-0.8. After the DNA dry powder is fully oscillated and dissolved with a certain volume of water, the OD value is measured by diluting with 1 mL of water. OD value of mother liquor should be converted according to dilution ratio. For example, to verify whether the amount of 2 OD primer is accurate, the simple practice is to add 1 mL water, thoroughly dissolve and mix, take 100 μL, add 900 μL water, a quartz colorimetric cup with a light diameter of 1 cm, a wavelength of 260 nm, and the absorbed reading at this time is 0.2.

This situation is more likely to happen when the Oligo DNA is shorter, and the difference between long-stranded Oligo DNA is smaller. There are two main reasons.

(1) different components of A, G, C and T and different electrophoresis speeds.

(2) The steric structure of DNA is different and the electrophoresis speed is different.

The amount of double-stranded DNA (e.g. plasmid DNA) can usually be judged by EB staining, because EB stains nucleic acids by embedding them between the double helix. Synthetic single-stranded DNA, on the other hand, can only be stained by EB by forming a local hairpin loop structure by folding back on itself or by forming a partial double helix structure between the strands. Due to the different base composition, the possibility of forming secondary structure differs from primer to primer, and the degree of EB staining will also vary, for example, Oligo(dT), etc. does not form secondary structure, and the EB staining effect is very poor. Therefore, EB staining cannot be used for quantification, and UV spectrophotometry should be applied for detection.

The longer the primer, the higher the probability of problems. Unless there is a special need, we recommend not to synthesize fragments longer than 80 mer. According to the current primer synthesis efficiency, the percentage of full-length primers will not exceed 40% for a crude product of 80 mer, and a lot will be lost in subsequent processing, so the final yield is very low. The maximum length of primers that can be synthesized is 145 bases.

It is important to use denaturing PAGE electrophoresis for primers. Since the primers are single-stranded DNA, it is easy to form a complex steric structure, so when Agarose electrophoresis is performed, it is easy to have multiple bands or no bands, and it is even more impossible to quantify by Agarose electrophoresis.

(1) Stability and Efficiency: Modified monomers are less stable and have longer coupling times, resulting in lower efficiency and yield.

(2) Purification: They require intensive purification methods like PAGE or HPLC, leading to significant material losses.

(3) Raw Material Cost: The raw materials for modified primers are much more expensive, often hundreds of times more than those for general primers.

There are several modifications that can block 3' extension. Most 3' modifications will block extension during PCR.

The modifications most frequently used for this purpose are the 3'ddC, 3' Inverted dT, 3' C3 spacer, 3' amino, and 3' phosphorylation.

We do not recommend PAGE purification for dye labeled oligos due to the presence of ammonium persulfate, a component of the PAGE gel. Ammonium persulfate can break linkages and may damage the ring structures that comprise many fluorescent dyes.

PAGE gels are exposed to UV light when the oligos are excised from the gel. UV light exposure can also damage fluorescent dyes. HPLC purification provides a pure product without damaging fluorescent modifications.

First check the baseline-corrected or ROX-corrected raw curves. Low signal values in the amplification curves are mostly due to high background fluorescence. In probe-based detection, high background fluorescence is often caused by poor probe design, leading to insufficient quenching by the quencher group, improper pairing of the reporter and quencher groups, or low fluorescence labeling efficiency of the probe.

Erratic peaks in melting curves can result from contamination in the reaction system. Confirm contamination using NTC and NRC results, and check potential sources such as water, oligos, enzymes, and the experiment settings. Reagents exposed to strong light or high temperatures may degrade, so it is recommended to compare results with fresh reagents.

If the water in which you dissolve the primers has a low pH or is contaminated with bacteria or nucleases, the primers will degrade. Inadequate thawing and mixing during use, and uneven liquid may also cause inaccurate amounts of primers. It is recommended to aliquot primers, avoid repeated freezing and thawing, and use 10 mM Tris pH 7.5 buffer to dissolve primers. There is another possibility that there is no problem with the primers, but that the quality of the materials used in PCR, especially the template, is not completely consistent with that used previously.

Basically not. Nowadays, various PCR amplification techniques and high-temperature polymerases have been developed to solve the problems of amplification failure and low amplification efficiency encountered in PCR amplification. For example, slot PCR amplifies gene fragments with very low copy number. Some repeat fragments, fragments with high GC content amplification, must use special amplification means to amplify.

Amplification does not come out, mainly the following two situations are more common.

(1) RT-PCR. many genes are difficult to amplify by conventional RT-PCR methods. the key to successful RT-PCR lies in the quality of the RNA of the RT reaction and the content of the target gene in the particular tissue and cell.

(2) Amplification from the genome. Generally, genes are single copies in the genome, and the genome needs to be used as a template in strictly controlled amounts. Too much genomic DNA can affect the Mg2+ concentration and pH in the reaction system.

No. We have analyzed some non-specific bands and sequencing found that at least one primer sequence can be found at both ends of these non-specific fragments. Therefore non-specific amplification is usually the result of template contamination (e.g. contaminated genome in RNA) or unsuitable amplification conditions.

Tsingke ensures sample uniformity by isolating monoclonal clones. Each clone is validated by Sanger sequencing, covering the entire gene and at least 50 bp of the vector backbone upstream and downstream, which ensures the gene is correct and properly placed in the vector. The plasmid then undergoes NGS to verify the accuracy of the entire plasmid and detect any contamination. Before shipment, a strict QC review is conducted.

Cancer research probes, basic research probes, hematologic probes, pathogenic probes, genetic disease probes, methylation probes.

1.Primer Length: Primers should be 18-24 bases long to minimize the likelihood of primer dimer formation.

2.Tm Values: The melting temperatures (Tm) of all primers in the multiplex reaction should be similar to ensure uniform annealing.

3.3'-Nucleotide Complementarity: Avoid complementarity at the 3' ends of primers to prevent the formation of primer dimers.

4.Individual Optimization: Optimize each pair of primers independently before pooling them. Once pooled, mix and optimize them sequentially.

The optimal annealing temperature for each primer pair needs to be determined individually, and the lowest Tm value among the primers should be used for efficient annealing in the multiplex reaction. Additionally, using the minimum number of amplification cycles necessary to achieve sufficient product yield helps ensure the efficiency and specificity of the PCR.

In multiplex PCR, multiple templates are amplified simultaneously, which can make enzyme and nucleotide concentrations limiting factors. Additionally, the time required to fully synthesize all products increases.  Multiplex PCR requires longer extension times compared to single-target PCR to ensure complete synthesis of all products.

Multiplex PCR primers should be 18-24 bases long. Longer primers are more likely to form primer dimers.

Primer synthesis is a multi-step chemical reaction, and the efficiency of each step of synthesis is 99% at most, and by-products cannot be avoided. The longer the chain, the higher the cumulative frequency of mutations. When you clone and sequence after PCR amplification, in order to save time and improve the success rate, we have the following suggestions.

(1) Please prepare 2~3 positive clones of the bacteriophage after you detect the positive clone, and try to send 2 or more clones for sequencing, so that the success rate will be greatly improved and a lot of time will be saved.

(2) You can also send one clone for testing first, and store the rest two clones of the bacteriophage in the refrigerator at 4 degrees, and send the remaining two clones for testing as soon as individual point mutations or deletions occur.

(3) This will have a very high probability of getting the correct sequence and will eliminate the need for a series of experimental operations such as re-PCR, ligation, cloning and screening, saving even more time.

If you find more than 2~3 clones with mutations in the primer region, and it is confirmed that it is caused by the primer, we will arrange for the expedited free recombination immediately and send it to you as soon as possible.

We specialize in synthesizing RNA oligos up to 180 nucleotides in length, and the types mainly include siRNA, sgRNA, miRNA, ASO, and various modified RNAs.

All RNA oligos undergo rigorous quality control:

(1) Mass spectrometry (MS): Every batch is verified by MS to confirm identity and mass accuracy.

(2) HPLC purification: Available for >90% purity requirement applications.

(3) Nuclease-free and integrity checks: Gel electrophoresis and functional validation are optional upon request.

We guarantee sequence accuracy, modification integrity, and high batch-to-batch reproducibility.

The entire process—from synthesis and QC to packaging—is highly customizable. In addition, we offer end-to-end solutions for small nucleic acid screening and delivery, including targeted delivery systems. Our services support high-throughput screening and provide comprehensive analyses such as mRNA/protein silencing efficiency, stability, off-target effects, immunogenicity, and cytotoxicity. The results are reliable and highly reproducible, helping you identify candidate molecules with superior silencing efficiency.

We offer a wide range of RNA modifications including:

(1) Stability enhancements: 2′-O-methyl (2′-OMe), 2′-fluoro (2′-F), phosphorothioate (PS) bonds

(2) Functional handles: 5′/3′ amino (NH2), thiol (SH), biotin, azide, DBCO

(3) Labels & detection: Fluorescent dyes (FAM, Cy3, Cy5), quenchers (BHQ)

(4) Specialty bases: Pseudouridine (Ψ), 5-methylcytidine

These modifications improve stability, specificity, delivery, and functionality for applications such as RNA therapeutics, diagnostics, and functional genomics.

A positive control is crucial for validating the experimental system. When the expected results from the RNA positive control are observed, it ensures that the transfection, RNA extraction, and QC methods used are reliable.

FAM has a maximum absorption at 495 nm and maximum emission at 520 nm. Cy3 has a maximum absorption at 550 nm and maximum emission at 565 nm. Cy5 has a maximum absorption at 643 nm and maximum emission at 667 nm.

(1) The product is shipped in the form of lyophilized powder, the dry powder is attached to the wall of the tube and is easily dispersed when opened, so please centrifuge the tube for a few seconds before dilution to make the product gather at the bottom of the tube and open the cap carefully, cover the tube after dissolution, vortex and shake to mix to make full dissolution. It is recommended to store at -20°C or below, avoid repeated freezing and thawing, and keep in separate containers. Centrifuge instantly before use and prepare 20~100 μM storage solution with RNase-free Water or ddH2O, the average molecular weight of siRNA is 13300.

(2) RNA handling rules should be strictly followed and RNase-free lab supplies should be used for experimental operations to avoid degradation of RNA. Please keep on ice when using this product.

(3) For RNA with fluorescent marker, we use brown centrifuge tube to pack, because the fluorescent marker is sensitive to light, it must be stored away from light.

Before opening, centrifuge the tube. Add sterile RNase-free H2O or ddH2O to prepare a 20 μM stock solution. Aliquot as needed and store at -20°C to -80°C.

Positive controls are verified siRNAs targeting housekeeping or reporter genes, used to assess the feasibility of the experimental methods. Tsingke provides effective siRNA positive controls targeting GAPDH, ACTB, GFP/EGFP.

Negative controls are non-specific siRNAs used to demonstrate the specificity of siRNA effects and can be selected from universal or scrambled sequences based on requirements.

Normally, the results of negative controls should be similar across different groups. Significant deviations may indicate issues with the accuracy of experimental results.

For certain genes, particularly those related to cellular stress or immune response, varying responses to external stress might cause inconsistent gene expression across control groups.

The two most common reasons for unsatisfactory silencing effects are low transfection efficiency and suboptimal siRNA sequence design.

(1) Check Transfection Efficiency: If you are using siRNA for the first time or working with a new cell line, we recommend testing and optimizing the transfection efficiency. If you have already optimized the transfection conditions and the problem persists, consider switching to a different transfection reagent or method, as this might improve transfection efficiency.

(2) Evaluate siRNA Sequence Design: If the transfection efficiency has been improved but the silencing effect is still not satisfactory, it may be due to ineffective siRNA sequence design.

Usually, FAM shows green fluorescence (excitation wavelength 495 nm) and CY3 shows red fluorescence (excitation wavelength 550 nm) after transfection. If no fluorescence is observed, consider the following:

(1) Is the fluorescence excitation wavelength correct?

- Ensure that the excitation wavelengths are set to 495 nm for FAM (green) and 550 nm for CY3 (red).

(2) Were the cells washed immediately after transfection?

- Wash the cells immediately after transfection to observe fluorescence. Delaying this step can result in reduced fluorescence.

(3) Was the siRNA product stored properly?

- Improper storage can lead to siRNA degradation and FAM fluorescence quenching. FAM is particularly sensitive to light, so store it away from light.

(4) Was the transfection performed away from light?

- Operate away from light during transfection. Avoid exposing FAM to white light for extended periods.

(5) Was fresh medium replaced in time after transfection?

- Prolonged transfection times without timely replacement of fresh medium can affect fluorescence. Ensure fresh medium is added promptly post-transfection.

Poor cell health before transfection can lead to increased cell death. Ensure cells are in optimal growth condition, as healthy cells better tolerate transfection reagents. High concentrations of siRNA or transfection reagents can be cytotoxic, so adjusting these concentrations may be necessary. Some transfection reagents are more toxic; consider switching to a less toxic alternative. Low-purity siRNA may contain impurities that affect cell viability; therefore, using high-purity siRNA is recommended. Additionally, prolonged transfection times can exacerbate cell death; ensure timely replacement of fresh medium after transfection.

Low efficiency of cell transfection can occur due to several reasons:

Insufficient siRNA purity may lead to RNA enzyme contamination. Use DEPC-treated consumables and reagents to prevent this issue.

Adjusting the concentration of the siRNA-transfection reagent complex can enhance transfection efficiency.

Serum in the transfection mixture can interfere with efficiency. Prepare the mixture without serum to minimize this effect.

Avoiding antibiotics in the culture medium is recommended as they can inhibit transfection efficiency.

siRNA-mediated RNA interference (RNAi) is a transient phenomenon and cannot be stably propagated through cell generations. Its effects typically do not last for an extended period, and it is generally recommended to complete detection within 3-4 days post-transfection. The optimal detection time varies depending on the cell type and target gene, but it is usually between 24-48 hours post-transfection. It is generally suggested to assess mRNA levels at 24-48 hours and protein levels at 48-72 hours post-transfection.

Transfection efficiency depends on the characteristics of the cells and the transfection method used, and it is not directly related to the siRNA sequence. Therefore, siRNA transfection efficiency may vary across different cell types.

Usually, the package products are guaranteed to interfere with about 70% of the target gene, if the interference effect is about 60%, we also consider the interference to be meaningful. If there is a significant change in the phenotype and function of the cells, it is also recognized in the literature. Generally, the interference efficiency can be improved by increasing the transfection efficiency and optimizing the experimental system.

siRNA acts directly on mRNA, so mRNA level detection is the most direct detection indicator. Many clients believe that the direct result of mRNA degradation should correspond to a decrease in protein content, and therefore protein level assay results can also be used as an indicator of effectiveness. In fact, in many cases there is often a mRNA decline level that does not correspond to the protein decline level. The possible reasons for this are:

Related to the time chosen for the assay. It may be that the decline in mRNA has not yet been reflected in changes in protein amounts or has not reached a level sufficient for detection, so it is generally recommended that protein and functional assays be pushed back slightly.

The translation process of mRNA is very complex, and the expression of genes in the cell must always maintain a balance, and after a certain level of expression of some proteins is sufficient to maintain their cellular function, the expression may be temporarily "turned off" and the transcribed Some mRNAs may not be involved in the process of protein translation, therefore, the down-regulation of mRNA level is not exactly positivelycorrelated with the down-regulation of protein level.

Different cells have different transfection efficiency and different gene expression levels, which are all related to the action efficiency of siRNA, so there is no guarantee that the interference effect is the same in different cells.

Purified guide RNAs support the same modifications as standard desalt purifications. These include standard guide RNA modifications and bases such as RNA, 2'-O-Methyl bases, and phosphorothioate bonds. They also include novel guide modifications such as 2' Fluoro and DNA bases, subject to complexity limitations.

Chemically synthesized sgRNA offers the following key advantages:

(1) Higher editing efficiency: A lot of studies have demonstrated that chemically synthesized sgRNA achieves higher editing efficiency compared to IVT sgRNA.

(2) Lower cytotoxicity: IVT sgRNA carries a 5′-triphosphate modification, which can trigger innate immune responses and lead to cytotoxicity, thereby interfering with experimental results. In contrast, chemically synthesized sgRNA avoids such off-target immunostimulatory effects.

(3) Enhanced stability: Chemical synthesis allows the incorporation of stabilizing modifications such as phosphorothioate (PS) bonds and methoxy groups, improving stability during storage and experimental procedures. Moreover, the chemical synthesis process ensures higher batch-to-batch consistency and reproducibility.

(4) Ease of use: Chemically synthesized sgRNA is ready to use upon delivery, while IVT sgRNA requires 2–3 days and 5–6 steps for production, incurring higher economic and time costs.

Using synthetic sgRNA eliminates the need for annealing crRNA and tracrRNA before application. More importantly, several studies have shown that long single-stranded sgRNA exhibits greater stability and higher editing efficiency compared to the crRNA:tracrRNA duplex when complexed with Cas9.

Phosphorothioate (PS) bonds are added to antisense oligonucleotides to protect them from nuclease degradation.  Antisense oligos can include a full PS backbone;  however, the Tm decreases with each PS bond added.  Including extensive PS modification can also promote the antisense oligo to non-specifically bind to proteins.  This characteristic can increase oligo circulation time and improve cellular internalization when a delivery tool is not employed.  However, it can also cause a pro-inflammatory response if the antisense oligo binds non-specifically to immune receptors.

The recommended initial transfection concentration for ASO is 50 nM, with detection times of 24-72 hours. Optimal concentration typically falls between 10-100 nM and should be determined through gradient screening.

Increase transfection efficiency: Optimize transfection conditions, cell type, and transfection reagents.

Optimize ASO concentration: Design a wide range of ASO concentration gradients to test and find the most suitable one.

Change the ASO sequence: Consider alternative sequences if secondary structures at the target site limit effectiveness.

There are several different ways to introduce oligos into cells, including use of liposomes, attaching the oligo to a ligand (e.g. peptides, GalNAc, cholesterol, fatty acids, aptamers, antibodies etc.), electroporation, and gymnotic (naked) delivery.

The best method depends on the type of cells and types of modifications on the oligo, as well as the intended use of the ASO. Contact us for more information.

The common purification methods are PAGE and HPLC.

For the modifications tested at this point, the pattern of degradation seen for modified oligos is similar to that for standard oligos. However, stability or shelf-life information has not been collected for all modifications offered.

For ideal stability, oligos that are to be stored long term should be stored frozen, at –20°C. If the oligos are going to be resuspended, storage in a TE buffer (10 mM Tris, 0.1 mM EDTA, pH 7.5–8.0; such as IDTE) is better than using water (non-DEPC treated). The impact from the type of storage medium used is marginal at  –20°C but becomes more important at higher temperatures. It is ideal to store oligos in the dark. Exposure to UV light should be avoided, and ambient lab light should be minimized, particularly for some types of modified oligos.

(1) Stability and Efficiency: Modified monomers are less stable and have longer coupling times, resulting in lower efficiency and yield.

(2) Purification: They require intensive purification methods like PAGE or HPLC, leading to significant material losses.

(3) Raw Material Cost: The raw materials for modified primers are much more expensive, often hundreds of times more than those for general primers.

FAM

TET

HEX

TAMRA

ROX

Cy3

Cy5

With fluorophores, it's important to remember that exposure to natural light or ozone can negatively impact stability. If fluorescently modified oligos will be exposed to sustained light, protective cover from the lighting source (e.g., using brown tubes and/or foil covering) should be considered.

Dual-labeled TaqMan® probes incorporating quenchers such as TAMRA, Eclipse, or BHQ are widely used in real-time PCR for accurate detection. The key differences lie in their fluorescence emission properties and spectral compatibility:

TAMRA is a fluorescent quencher—it emits background signal itself, which may reduce sensitivity. Eclipse & BHQ are non-fluorescent quenchers (NFQs), offering lower background and higher signal-to-noise ratio.

For multiplex qPCR:

(1) Eclipse has a broad quenching range (390–625 nm), supporting common dyes like FAM, HEX, TAMRA, and ROX.

(2) BHQ series cover an even wider spectrum (430 nm to near-infrared), making them ideal for multiplex assays with dyes such as Cy3 and Cy5.

For high-sensitivity and multiplex applications, NFQ-based probes (Eclipse or BHQ) are strongly recommended.

Amino modifiers introduce a reactive amino group (−NH2) to oligonucleotides, enabling covalent conjugation with various molecules such as fluorophores, biotin, or surfaces.

Key points:

(1) The 5'Aminolinker (C6) modifier is attached to the 5′ sugar ring via phosphoramidite chemistry during synthesis (not to the terminal base).

(2) The amino group absorbs at 210 nm (not 260 nm) and cannot be detected by standard agarose or acrylamide gel electrophoresis.

(3) The 3'Aminolinker(C7) is compatible only with certain 5′ modifications (e.g., FAM, HEX, TET, Fluorescein, Biotin, phosphate). It is incompatible with dyes like Alexa Fluor® that also require an amine for conjugation.

(4) Amino modification remains a simple and cost-effective strategy. For most applications, the 5'C6-NH2 Modification is highly recommended.

Biotin-modified oligonucleotides leverage the strong biotin-streptavidin interaction, enabling diverse applications in research and diagnostics. Key uses include:

Non-radioactive immunoassays for protein detection

Intracellular chemical staining and cell separation

Nucleic acid isolation and hybridization to detect specific DNA/RNA sequences

Probes for studying conformational changes (e.g., in ion channels)

Immobilization on solid-phase surfaces (e.g., beads, chips)

For conjugation to a gold electrode, mix 1 µL of 1 mM TCEP with 99 µL of thiol-modified DNA (at a concentration of 0.2 µM) and incubate at room temperature for 30–60 minutes. The reduced DNA may be used directly in subsequent experiments without removing TCEP. If a higher DNA concentration is required (e.g., 100 µM), 10 mM TCEP should be used for reduction. In this case, the amount of TCEP must be at least 100 times the molar amount of DNA. For conjugation to gold nanoparticles, mix 200 µL of 15 µM thiol-modified DNA with 5 µL of 1 M TCEP and reduce at room temperature for 30 minutes. The mixture can then be directly added to the gold nanoparticle solution.

Common stabilizing modifications include:

(1) 2′-O-Methyl (2′-OMe): Enhances nuclease resistance

(2) 2′-Fluoro (2′-F): Increases binding affinity and stability

(3) Phosphorothioate backbone: Improves resistance to enzymatic degradation

(4) Locked Nucleic Acid (LNA):“Locks” the structure for extreme stability and high affinity

They enable the development of advanced therapeutics by:

(1) Enhancing target engagement and specificity (e.g., LNA, cEt)

(2) Improving pharmacokinetics (e.g., GalNAc for liver targeting)

(3) Reducing immunogenicity (e.g., 2′-OMe, pseudouridine)

(4) Facilitating efficient delivery (e.g., cholesterol conjugation)

The choice depends on your goal:

For stability: Choose 2′-OMe, 2′-F, or LNA

For delivery: Consider GalNAc or cholesterol

For epigenetics: Use 5-methyl-dC or 5hmdC

For high-affinity binding: LNA or cEt are ideal

For functional conjugation: Biotin, amines, or thiols may be used

These are chemical alterations made to the natural sugar-phosphate backbone of oligonucleotides. They are designed to enhance properties such as nuclease resistance, stability in biological environments, and the ability to attach other molecules or enable site-specific cleavage.

Synthesized oligos have a hydroxyl group at the 5' end and are not phosphorylated.

If 5' phosphorylation is required, you can phosphorylate them using polynucleotide kinase, or request phosphorylation at the 5' or 3' end when palce your order.

Several modifications enable conjugation:

(1) Maleimide: For covalent attachment to thiol (-SH) groups on proteins.

(2) Aldehyde or Carboxy: For reaction with amine (-NH₂) groups on surfaces or proteins.

(3) DBCO/Azide: For highly specific, biorthogonal click chemistry.

Spacer-modified primers should be stored under the following conditions to ensure stability and prevent degradation:

(1) Protect from light: Store in light-blocking or opaque containers, as they are sensitive to light-induced degradation.

(2) Low-temperature storage: Keep frozen at –20°C or below in a refrigerator or freezer to minimize oxidation and decomposition.

(3) Dry conditions: Store in a sealed container with desiccant to avoid moisture and oxygen exposure.

(4) Avoid freeze-thaw cycles: Aliquot upon receipt to prevent repeated freezing and thawing, which reduces bioactivity and chemical stability.

The dSpacer is a unique modification that mimics a natural nucleotide missing its base (an abasic site). Its primary application is in structural studies to investigate how enzymes (like those in DNA repair pathways) recognize and process damaged DNA. It can also be used to introduce flexibility into an oligonucleotide structure.

The choice depends on the required length and application:

(1) Spacer C3: A short, 3-carbon chain.  Often used as a minimalist spacer or to block the 3'-end of an oligo to prevent enzymatic extension.

(2) Spacer C6: A versatile 6-carbon chain, very common for attaching molecules like fluorophores or biotin.

(3) Spacer C12: A longer, 12-carbon chain used when greater separation between molecules is needed.

We require the DNA or amino acid sequence you need synthesized.If codon optimization is required, specify the host species. Additionally, indicate any required restriction sites at the 5' and/or 3' ends, any internal modifications to avoid, and if you need additional sequences added (e.g., Kozak sequence, stop codons, etc.).Or you can download the "Tsingke Gene Synthesis Order Form" from our website and email it to info@tsingke.com.cn.Our technical support team will promotly estimate the pricing and turnaround time for you and send you a quotation shortly.

To help us design the most effective shRNA, please provide:

Gene information: Official gene name (Gene Symbol), species or Gene ID, and the target sequence.

Vector preference: The vector you would like the shRNA sequence to be cloned into. You may select from our available vectors (based on selectable markers, etc.) or provide your own vector.

Or you can download the "Tsingke shRNA vector construction Order Form" from our website and email it to info@tsingke.com.cn. Our technical support team will promotly estimate the pricing and turnaround time for you and send you a quotation shortly.

Download the "Tsingke Gene Synthesis Order Form" from our website, complete the “Gene info”, “Cloning vector”and“Plasmid preparation”section, and send it via email to info@tsingke.com.cn to place your order.

Yes, all customers' information will be kept confidential.

Yes, Tsingke also provides protein expression and purification services for synthesized genes, offering six major protein expression systems: E. coli protein expression system, Bacillus subtilis protein expression system, yeast protein expression system, insect cell protein expression system, mammalian cell protein expression system, and prokaryotic cell-free protein expression system.

Yes, Tsingke offers free codon optimization.

Codon optimization enhances gene expression by using preferred codons and avoiding rare ones for a specified host. This optimization correlates strongly with improved gene expression levels. Tsingkes unique Geneoptimizer software supports optimization for tens of thousands of hosts and offers customized services. Key optimization parameters include codonpreference. GC content, restriction enzyme sites (deletion), poly structures, and sequence repeats.

By default, we provide pUC57 vectors containing Amp (ampicilin) for free. we also offer a library of over 200 vector options available for free.

Generally, the default clone is between EcoRⅠ and Hind Ⅲ. If you have particular needs, please note and we can select other suitable cloning sites according to the experimental needs based on your sequence.

Yes, you can use your own vector. it is recommended to ship your vector in advance; otherwise, the turnaround time may be extended by 5 calendar days to verify the vector.

We need you to ship at least 2 μg of the plasmid and provide the full sequence information of the vector.lf the full sequence is not available, please provide at least 200 bp upstream and downstream of the cloning site, along with the vector's antibiotic resistance information.

No, the order defaults to providing 4 μg of plasmid dry powder. This plasmid is not recommended for direct transfection into cells or protein expression. If you need to conduct related experiments, please perform plasmid transformation, then isolate a single colony for cultivation, and use a plasmid extraction kit that meets the required standards to extract high-quality plasmid. Our company also provides plasmid extraction services. If you have higher requirements for the plasmid, you can select our plasmid extraction service simultaneously when placing your order.

The synthesized plasmid can be stored for one vear. Customer-provided vectors that are free of issues will also be stored forone year. During the storage period, you can use the plasmid for site-directed mutagenesis, cloning, or other services.

If there are any quality issues within one month of receipt,we will provide a free replacement. Overseas shipping fees will be charged separately, or the plasmids can be sent with other orders.

High and low GC content, repetitive sequence:

Solution: Divide the sequence into small segments to reduce the impact of structure, and choose the optimal connectionsolution, such as enzyme digestion and ligation and multi-segment recombination.

Long sequences

Solution: use multi-segment recombination to construct divided gene fragments, and combine them with vitro and vivo assembly.

Special parts of vectors (e.g. ccdB, LTR)

Solution: Use different competent cells corresponding to different parts of vectors, such as DB3.1, which renders the strain resistant to the toxic effects of the ccdB gene, the recombination-deficient Stbl series, etc.

Other unpredictable dlificulties, such as genetic instability and toxicity, and the introduction of random mutations and deletions.

Solution: Try different competent cells.

Tsingke ensures sample uniformity by isolating monoconal clones, Each clone is validated by Sanger sequencing, covering the entiregene and at least 50 bp of the vector backbone upstream and downstream, which ensures the gene is correct and properly placed inthe vector. The plasmid then undergoes NGS to verify the accuracy of the entire plasmid and detect any contamination. Beforeshipment, a strict OC review is conducted.

A tube containing approximately 500 ng of lyophilized DNA, lectronic Certificate of Analysis (COA), sequencing chromatograms (.abi files) and Target Sequences (.seq files)if sequencing validation is needed.

We provide one tube of lyophilized plasmid (stored at -20°C).

Our company defines long DNA sequences as those exceeding 10 kb. For sequences ranging from 10 kb to 30 kb, yeastassemoly methods are usedto first construct them into the pcc1413 vector before transferring them to your specified targetvector. sequences exceeding 30 kb will be directly constructed into the pcc1413 vector.

Efficient construction: Rapid generation of shRNA expression plasmids via gene synthesis, eliminating tedious cloning steps.

Professional optimization: Customized design for any gene from any species, with sequence optimization to enhance knockdown efficiency and minimize off-target effects.

Versatile vectors: A wide range of lentiviral backbones available; shRNA can be delivered by direct transfection or packaged into lentiviral particles for infection.

Easy selection: Vectors carry common resistance markers (e.g., PURO/NEO) for stable cell line establishment; fluorescent markers (e.g.,GFP/mcherry) allow real-time monitoring of transfection/infection efficiency and enable flow cytometry sorting.

Yes, Tsingke offer end-to-end services from plasmid to ready-to-use viral particles, including packaging, concentration, and titer determination. Lentivirus, AAV, and adenovirus systems are available, delivering high-titer viral particles ready for your experiments.

Download the "Tsingke Gene Synthesis Order Form" from our website, fill in the relevant fields according to your needs, and send it via email to info@tsingke.com.cn to place your order.

a. Templates: Provide and send us the template plasmid(≥2μg). If the plasmid was constructed by Tsingke and the order was placed within the last year, you do not need to provide it again.

b. Vector details: Choose from our 200+ free vectors or provide your vector.

c. Provide the sequences and resistance information for both template and vector, along with the target sequence and cloning siteinformation.

d. Order Form: Download the "Tsingke Gene Synthesis Order Form" from our website, fill in the relevant fields according to your needs,and send it via email to info@tsingke.com.cn to place your order.

No, the template sequence needs to be first constructed into a T-vector or another vector. then, amplify using the plasmid asthe template. Alternatively, you can choose our gene synthesis service by simply providing the DNA sequence.

a.DNA template: Please send us your template

b.Information: The sequence of the template including the target gene and vector information, mutation specifications.

c.Order Form: Download the "Tsingke Gene Synthesis Order Form" from our website, fillin the relevant fields according to your needsand send it via email to info@tsingke.com.cn to place your order.

The pricing for site-directed mutagenesis is evaluated and quoted based on the specific location of the mutation sites and the numberof mutations (all mutations within a 30-base region will be considered as one mutation).

Tsingke provides high-quality plasmid extraction services with Research Grade and Transfection Grade to meet diverse experimental needs.

We provide the following two options:

a. Gene synthesis + plasmid preparation: Synthesize your gene by Tsinkge our and then proceed with plasmid preparation.

b. Plasmid preparation only: You need to ship the target plasmid to us.

Download the "Tsingke Gene Synthesis Order Form" from our website, fillin the relevant fields according to your needs, and send it via email to info@tsingke.com.cn or gene@tsingke.com.cn to place your order.

Yes, Tsingke provides transfection-grade plasmid preparation services with low endotoxin levels and high supercoiled DNAcontent, suitable for cell transfection, gene therapy, and gene editing. Endotoxin levels can be selected as< 0.1 Eu/μg, <0.01 EU/μg, or <0.005 EU/μg.

Tsingke offers an integrated solution from gene synthesis to plasmid preparation, ensuring high-quality plasmids delivered in a seamless and efficient workflow. This one-stop service saves time, reduces experimental complexity, and guarantees accuracy for your downstream applications.

Adeno-associated virus(AAV):Suitable for long-term gene expression and applications with high safety requirements (e.g., gene therapy).

Advantages: Long-term expression, high safety.

Disadvantages: Lower transduction efficiency, small gene capacity.

Lentivirus(LV):Suitable for stable transduction and long-term gene expression (e.g., cell therapy).

Advantages: High transduction efficiency, broad cell type applicability, long-term expression.

Disadvantages: Potential genome integration, safety risks.

Adenovirus (ADV):Suitable for short-term expression and large-scale cell transduction (e.g., vaccine development).

Advantages: High transduction efficiency, suitable for large-scale production.

Disadvantages: Short-term expression, can trigger immune response.

Choose the appropriate vector based on your experimental needs.

Common methods for gene overexpression:

Overexpression plasmids: Introduce the target gene into cells through plasmid transfection for overexpression.

Overexpression lentivirus: Use lentiviral vectors to stably transduce the target gene, suitable for long-term gene expression.

CRISPRa transcriptional activation system: Utilize CRISPR technology to activate the expression of endogenous genes, offering precise regulation.

Each method has its own characteristics, and the choice should be based on the experimental objectives and cell types.

Turnaround time varies by vector type and project scale. Typically, AAV, LV, and AdV vectors can be delivered in as fast as 12–18 business days. Larger-scale or customized projects may take longer.

No. Our service includes plasmid preparation, where we clone the gene or sgRNA cassette into an appropriate transfer plasmid and prepare high-quality DNA.

Yes. For each category of viruses (AAV, ADV, lentivirus, etc.), clients can opt for a guarantee service to ensure the viral vectors meet expected performance standards.

Yes. We offer custom stable cell line construction, including knockout, overexpression/interference, point mutation, and knock-in cell lines.

• Delivered as frozen or cultured cells (≥1×10⁶ cells per vial)

• Turnaround time: 6–11 weeks depending on cell type

• Quality verification: Sanger sequencing, qPCR, or protein-level validation

• Includes positive clones and, when applicable, control lines ready for downstream experiments

The GFP fluorescence intensity in target cells depends on several factors, including the number of viral particles transducing the cells, the proliferative state of the cells, the cell type, and the observation time. Generally, the higher the number of viral particles transducing the target cells, and the faster the cell proliferation, the stronger the GFP fluorescence. For cells with faster proliferation, GFP expression typically peaks 48 to 72 hours after viral transduction. For slower proliferating cells, the GFP expression time will be extended.

At Tsingke, we generally improve transduction efficiency by increasing the MOI (multiplicity of infection). Additionally, maintaining the target cells in good growth condition is crucial to ensure normal transduction efficiency.

rAAV can package approximately 3.0 kb of genetic material. However, if you want to add a marker, such as GFP or RFP, the size of the gene that can be inserted will be smaller, around 1.5 kb.

Using your existing plasmid: The virus can be packaged and delivered in approximately 12–15 days.

Including plasmid construction: If we also need to construct the plasmid for your gene of interest, the full process typically takes 15–25 days, depending on the gene size, complexity, and any special design requirements.

Factors that can affect timing:

• Gene length (genes over 2 kb may require extra time)

• Sequence complexity or modifications

Additional quality control steps requested by the customer

（1）The most likely reason is: When lentivirus is used, an excessive viral dose can severely damage the cell genome and cell membrane. Therefore, it is necessary to determine the optimal MOI (multiplicity of infection) for each cell type, count the cells based on this value, and then use the appropriate lentiviral dose for the target cell infection.

（2）Other possible reasons: Lentiviral infection itself may induce cell differentiation, which affects the cell's proliferative ability.

（3）If the control lentiviral infection produces normal results but the target lentivirus shows abnormal results, it is possible that the gene targeted by the lentivirus is involved in processes such as the cell cycle, cell proliferation, cell metabolism, or cell adhesion. This situation is difficult to avoid but can generally be addressed by reducing the lentiviral dose, using an inducible expression system, or altering culture conditions.

（1）Functional effectiveness means: Validation is carried out on model cells (e.g., 293T, 3T3 cells) to confirm effectiveness.

（2）For overexpression and related lentiviruses, the expression of the promoter varies across different cell types. As a result, the same lentivirus infecting different cells may produce different effects.

（3）For RNA interference lentiviruses, the background expression levels of the target gene and the differences in the intracellular environment across different cell types may lead to different effects from the same lentivirus infecting different cells.

Our service provides high-titer, high-purity adenoviral vectors customized to your needs. Each delivery includes target and control viruses, precise titers, a detailed shipping list, and usage instructions for seamless integration into your experiments.

Adenovirus enables rapid, strong, and transient gene expression in a broad range of mammalian cells. It remains episomal without integrating into the host genome, supports large genes, and provides high transduction efficiency, making it ideal for in vitro and in vivo studies.

Viral Particles (VP) represent the total number of both live and dead viral particles and do not reflect the amount of active virus in the preparation. Plaque Forming Units (PFU) represent the number of infectious or live viruses, providing a measure of the active virus present. Infectious Units (IFU) are equivalent to PFU. For most virus preparations, the VP/PFU ratio is 20:1 to 50:1. Using VP as a measurement can result in significant variations in the amount of actual live viruses present, whereas using IFU or PFU gives more consistent outcomes.

Key indicators for assessing the quality of a mutation library include:

- Diversity of mutations: The mutations in the library should cover a wide range of variations in the target gene, ensuring sufficient representativeness.

- Complexity of the library: This refers to the number of mutant variants in the library, ensuring the library size is large enough to cover possible mutation combinations.

- Accuracy of mutations: The accuracy of mutations in the library should be verified using high-throughput sequencing techniques (such as NGS) to avoid the introduction of erroneous mutations.

- Consider the target cell type, as some cells are more sensitive than others.

- Higher titers aren't always better—too high can be toxic or trigger immune responses.

- Start with small-scale tests to determine the optimal functional titer.

- Use qPCR or Western blotting to check gene expression changes.

- Functional assays (like reporter assays or phenotypic readouts) can confirm the library's effect.

- Sequencing can confirm the guide RNA or mutation was correctly introduced.

Polyclonal stable cell line (cell pool): This is obtained by directly selecting cells with drug resistance after transfection. It consists of multiple clones from different sources, with varying integration sites and expression levels of the exogenous gene. It has a short production cycle and low cost, suitable for initial screening and high-expression applications.

Monoclonal stable cell line: This is derived from a cell pool through methods such as limiting dilution, and consists of a single clone of cells with stable integration of the exogenous fragment. All cells have the same integration site, ensuring consistency and stability in expression. It is suitable for high-precision research and biopharmaceutical production. However, it has a longer production cycle and higher cost, and is ideal for cell line experiments requiring subcellular localization, difficult-to-infect cells, and low-expression applications.

Each project includes comprehensive validation such as sequencing results, qPCR data, and optional protein-level verification (e.g., Western blot), ensuring reliable and reproducible results.

Overexpression cell lines are faster and more cost-effective to construct, and they are suitable for studying gene function at the protein level. However, expression may be non-physiological due to strong promoters or random genomic integration. Knock-in cell lines, on the other hand, place the gene under its natural promoter, allowing physiologically relevant expression and more accurate functional studies. The trade-off is that knock-in projects are more complex and typically require longer timelines.

Optimize elution conditions by gradually lowering the pH from a high value; neutralize the purified sample promptly, or even neutralize while purifying; prepare and store the sample at low temperatures during the purification process; add protein protectants such as glycerol and Tween; when antibody concentration is high, dilute in time to avoid the risk of aggregation due to high concentration.

The verification can be done from the following three aspects:

First, at the molecular sequence level, after constructing the expression vector, the expression plasmid is sequenced for confirmation. Sequencing files can be used for verification.

Second, during the expression process, a comparison is made between induced and uninduced lysis buffers. Differences in protein bands can be detected by SDS-PAGE. The difference can be observed at the theoretical size of the target protein.

Lastly, Western blot (WB) validation can be performed using tag antibodies or antibodies specific to the target protein.

If protein verification is needed, protein coverage analysis can be conducted by comparing the theoretical sequence with the actual coverage ratio. However, the purity of the target protein must be greater than 90%. If the purity is lower, the coverage ratio may be affected. For lower purity samples, gel slicing can be performed for further analysis.

The advantage of eukaryotic expression systems over prokaryotic systems is that the protein activity is generally better. In theory, most proteins expressed in eukaryotic systems are active. However, the stability of different samples may vary, so the actual experimental results should be used as the basis for determining protein stability.

Advantages of CHO Cells:

• Can grow stably in chemically defined, serum-free suspension cultures.

• Well-characterized genome and generally safe with respect to human pathogenic viruses.

• Capable of human-like post-translational modifications (e.g., glycosylation).

• Amenable to genetic modification and stable cell line generation.

Advantages of HEK293 Cells:

• Faster growth rate compared to many other mammalian lines.

• High cell density and very high transfection efficiency.

• Produce post-translational modifications that closely resemble human protein structures.

Clients need to provide the gene sequence. Once confirmed, our team will design the optimal expression strategy for your project.

Pricing depends on gene length and protein difficulty. For non-guaranteed expression services, if expression is unsuccessful, 50% of the cost may be charged. Guaranteed expression services ensure successful production. The default delivery is 1 mg, with larger amounts available upon inquiry.

Clients need to provide the peptide sequence(s). For modified or structured peptides, specify the modifications or structural type required.

Our services are designed for rapid turnaround: e.g. recombinant antibody production in 2–5 weeks, hybridoma sequencing in 1–2 weeks.

All antibodies undergo rigorous QC such as SDS-PAGE (>95%), HPLC purity checks, and endotoxin testing (<1 EU/mg). Sequencing services include raw data, sequence verification, and optional NGS validation.

Yes. We support full-length IgG, Fab, VHH, and polyclonal antibodies, across multiple species (mouse, rabbit, camelid). Clients can provide their own antigens or request custom antigen design and preparation.

To begin a project, clients need to provide the DNA sequence of the target protein. Any specific requirements regarding expression system, protein quantity, or purity can also be communicated to help design the optimal expression strategy. Additional services, such as endotoxin removal, protein purification, or QC options, can be requested based on the project needs.

Clients receive purified protein (with SDS-PAGE >85% purity), expression plasmids, optional expression strains, and a COA report. Additional QC options, such as SEC-HPLC, endotoxin testing, Western blot verification, and protein coverage analysis, are also available upon request.

For certain services, if protein expression fails, clients may be charged 50% of the project cost to cover the expenses already incurred. Guaranteed expression services include multiple strategies to ensure success, minimizing the risk of failure.

We can synthesize peptides up to 30 amino acids for standard orders, with scales ranging from 1 mg to 1 g for lab-scale projects, and up to 1 kg or more for large-scale/bulk synthesis.

Peptides can be provided at purity levels from 75% to 98%, allowing you to select the optimal balance between cost and experimental requirements.

Standard synthesis is optimized for peptides up to 30 amino acids. For longer sequences, complex modifications, or unusual amino acids, we recommend consulting with our team for customized solutions.

Depending on the service, some antibodies are provided as purified protein ready for use, while others, like plate supernatant antibodies, may require additional purification before sensitive applications.

The main differences are in quantity, turnaround time, and QC standards:

Standard Recombinant Antibody Expression: Designed for research-scale production (3 mL – >1 L). It has a fixed 2-week turnaround and high QC standards (SDS-PAGE >95%, HPLC >95%, Endotoxin <1 EU/mg). This service is ideal when the primary goal is rapid, small- to medium-scale antibody production.

Guaranteed Recombinant Antibody Expression: Provides a guaranteed yield (10–1000 mg) and a slightly longer turnaround (2–5 weeks). QC standards are also high (SDS-PAGE >95%, HPLC >90%, Endotoxin <1 EU/mg), but this service is specifically suitable when a precise amount of antibody is required for experiments or downstream applications.

Yes, we offer custom bispecific antibodies and tagged formats (e.g., VHH-His, VHH-Fc). Secondary purification can be requested to meet specific research requirements.

Customers should provide more than 1 × 10⁶ frozen hybridoma cells or Trizol-processed cells. Antibody subtype information and hybridoma species/source details are also needed.

The turnaround time is approximately 1–2 weeks, depending on the service type (variable region sequencing, full-length sequencing, or NGS high-throughput sequencing).

You will receive an electronic report containing the antibody light and heavy chain sequences, along with raw sequencing data (NGS in FASTQ format, no chromatogram files).

Our Antibody Screening Service covers monoclonal antibodies from single B cells (mouse and rabbit), as well as nanobodies from natural human or camelid libraries. Clients receive validated sequences and, where applicable, recombinant antibody proteins suitable for downstream research or therapeutic applications.

After supernatant screening, approximately 100 μL of culture supernatant from each positive clone is provided to the client for functional validation. Only the candidates that pass this validation proceed to antibody gene sequencing and recombinant expression.

Human antibody library: Derived from 100 donors, with a titer of 2×10¹³ PFU.

Camelid VHH (single-domain) library: Derived from 200 camels, with a titer of 2×10¹³ PFU.

Our Antibody Discovery service covers the full workflow from antigen design and synthesis, immunization, hybridoma or polyclonal screening, antibody purification, functional testing, and optional labeling or fragmentation. We provide high-affinity, validated antibodies ready for research or therapeutic applications.

The Rapid Rabbit service uses an optimized high-efficiency immunization system, reducing the immunization period to 4 weeks while still producing high-affinity, validated antibodies. This allows delivery of purified antibodies and functional data in just 7 weeks, compared to 9–11 weeks for standard polyclonal services, without compromising quality or specificity.

Yes! All antibodies are validated, and accompanied by ELISA or QC data, ensuring they are ready for research.