PCR involves the enzymatic amplification of nucleic acid templates, and can be divided into four major steps, listed below. 1 Denaturation: Double-stranded DNA separates into single strands. Annealing ...

Pipet the emulsion into the wells of a PCR plate as 10 aliquots of 50 μl. Pipet 50 μl of the aqueous phase (Step 3) into a well as a nonemulsified control. Overlay the emulsified and ...

If the template DNA is complex, such as mammalian cellular DNA, then we generally use 3–30 ng of template DNA per PCR (1,000–10,000 haploid genome equivalents). All steps are performed at room ...