In: Biology
Can DNA be amplified from a degraded sample for PCR?
Degradation of sample indicates that the DNA present in the sample is broken down to single nucleotides. DNA is degraded in samples by nucleases present in the sample.
Degraded sample can be amplified by PCR only if there is still some amount of intact template present in the sample. Intact template means that these DNA fragments have both the regions that are complementary to the forward and reverse primers. In this case, minimal amplification of the template will occur. The PCR product seen in agarose gel electrophoresis will be reduced significantly. However, if the sample is completely degraded, then no amplification will occur upon PCR.
Amplification of completely degraded sample in PCR will appear as a smear upon agarose gel electrophoresis. If only single stranded cuts are present in the degraded DNA, then smearing may not be observed. In this case, DNA samples need to be denatured before gel electrophoresis to check for degradation.