Frequently Asked Questions

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.