Custom Oligo Pools
● The error rate is only 0.5%, that is, only one error base in 200 bases is found.
● Each primer has a full-length DNA fragment of 1 fmole.
● The primers up to 170 bases can be synthesized.
● Can be used in synthetic biology, gene synthesis, targeted sequencing, and complex DNA library.
12K Oligo Pools (Up to 12,472 Oligos)
- 2-3 weeks turnaround time.
92K Oligo Pools (Up to 91,776 Oligos)
- 2-3 weeks turnaround time.
8M Oligo Pools (Up to 180,000 Oligos) are also available by inquiry.
Oligo Pools Application Guide
We can provide some common guidelines for primer design and amplification of oligo pools. But for specific experiments, our guidelines can only be used as general guidelines. You need to adjust the specific experimental steps according to the conditions. The guiding principles for the use of oligo pools mainly include the following points:
- A set of universal primers will be used before amplification. PCR amplification can achieve the effect of purifying oligo pools because the failed primer sequence (the primer sequence missing at 5 'end) cannot be amplified.
- To achieve a higher annealing temperature, we suggest using a high Tm value or longer primers. Higher annealing temperatures can reduce primer mismatch and enhance the specificity of amplification.
- In PCR amplification, we recommend not using conventional Taq polymerase, but using hot start enzymes, such as Fusion, Q5, Pfu, Vent, Platinum Taq Polymerase, and PrimeTaq™ HotStart Direct PCR DNA Polymerase. The selection of enzymes should be cautious, and some enzymes need to be tested before use.
- Due to the different sequence lengths, primer designs, and polymerase systems of different users, the usage of starting templates is also different. For PCR templates, it is suggested to use 1/100, 1/400, 1/1600, 1/6400, and 1/25600 of the total reaction volume for the gradient test. Set the number of PCR cycles, such as 25 cycles to amplify, and use gel or Bioanalyzer to detect the amount of PCR products. As long as the quantity of PCR products meets the requirements, the smaller amount should be chosen as the final amount of the template.
- If multiple sequences need to be amplified from the product, nested PCR is used to amplify all the sequences, and then internal primers are used to amplify each group of sequences. Only one round of PCR may not be enough for getting a good result.
- The basic strategy of primer removal depends on the specific needs of the downstream application of oligo pools, such as blunt end or sticky end.
- For normal cloning, users can insert the restriction sites recognized by type IIS restriction enzymes into the universal primers and then remove the universal primers after PCR amplification. These restriction enzymes can digest near the recognition site to maintain the integrity of variable sequences. After the digestion, the oligo sequences can be inserted into the vector.
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