Nested PCR (CAT#: STEM-MB-0196-WXH)

Nested PCR means that two pairs of PCR primers were used for a single locus. The first pair amplified the locus as seen in any PCR experiment. The second pair of primers (nested primers) bind within the first PCR product and produce a second PCR product that will be shorter than the first one. The logic behind this strategy is that if the wrong locus were amplified by mistake, the probability is very low that it would also be amplified a second time by a second pair of primers.

• Amplifying genes present in low abundance.
• Detection of microorganisms when they are present in very low quantities.

Step One: The DNA target template is bound by the first set of primers shown in blue. The primers may bind to alternative, similar primer binding sites which give multiple products however only one of these PCR products give the intended sequence (multiple products not shown).
Step Two: PCR products from the first PCR reaction are subjected to a second PCR run however with a second new set of primers shown in red.