Oligo Pool Resuspension and PCR
NUS iGEM
Published: 2023-10-09 DOI: 10.17504/protocols.io.8epv5xe3ng1b/v1
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Abstract
2023 NUS-Singapore iGEM team followed this protocol to resuspend the dried oligo pool and perform PCR to amplify the DNA fragments in the oligo pool.
Steps
Oligo Pool Resuspension
1.
Centrifuge the oligo pool for a few seconds before opening the tube.
2.
Calculate the volume of
3.
Add the respective volume of
Oligo Pool PCR
4.
Prepare a PCR tube, and add the following reagents:
| A | B |
|---|---|
| KOD OneTM PCR Master Mix (Blue) | 25μL |
| Each Primer (Two Primers In Total) | 1.5μL |
| Oligo Pool Sample | 1μL |
| DI Water | 21μL |
5.
Run a PCR with the following setting:
| A | B |
|---|---|
| 98°C | 10s |
| 60°C | 5s |
| 68°C | 10s |
| Go to step 1, repeat the cycle 10 times | |
| 68°C | 5 minutes |
| 12°C | Infinite Loop |
6.
Follow the PCR clean-up protocol to isolate the oligos from the PCR product and keep it at Room temperature .
PCR Clean-up
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