PCR Beginner's Guide: What if PCR does not expand again?

PCR Beginner's Guide: What if PCR does not expand again?

This problem is a common problem, that is, a PCR experiment has been done very well, and it will not work after stopping for a while. After a messy search for reasons, it still doesn't work, it is very painful!

Find the problem from big to small:

1. PCR amplification system problem
Using other ongoing and amplified normal templates and primers to prove that the PCR amplification system is no problem, ie PCR reaction mixture, water, liquid taker, PCR instrument, operation, etc. are all good (positive control)

2, the primer problem
Use the previously amplified product as a template (diluted 50 times) to prove that the primer has no problem.

3, just a template problem.
Because the probability of sample degradation is much higher than the possibility of primer degradation. It is also possible that there are too many inhibitors in the sample (not dissolved when fresh).

Don't rely on luck to find problems. It is important to do the first one first. Finding problems indiscriminately often makes people lose confidence.

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