Multiplex PCR Amplification of Ancient DNA

摘要: Multiplex PCR allows the simultaneous amplification of up to dozens target fragments in a single PCR. It is therefore powerful tool obtain many kilobases continuous sequence from minute amounts ancient DNA (aDNA), which usually must be amplified multiple short and overlapping fragments. Because significantly less template required compared amplifying each fragment separately, multiplex particularly beneficial when fossil material itself, or access material, limited. The recently refined two-step protocol consists first-step reaction (the actual PCR) that then acts as for second-step During second step, nested primers are used individual reactions. Although same set can both steps, using step adds an additional level selectivity specificity, minimizing artifacts. This important complex mixtures DNA, such aDNA extracts, amplified.