Frequently Asked Questions

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.