PCR Preamplification of cDNA

Get tips on using Power cDNA Synthesis Kit to perform cDNA synthesis Bacteria

Get tips on using qScript cDNA Synthesis Kit to perform cDNA synthesis Bacteria

Get tips on using iScript cDNA Synthesis Kit to perform cDNA synthesis Bacteria

Get tips on using iScript cDNA Synthesis Kit to perform cDNA synthesis Tissue

Get tips on using First-Strand cDNA Synthesis Kit to perform cDNA synthesis Yeast

Get tips on using First-Strand cDNA Synthesis Kit to perform cDNA synthesis Tissue

Get tips on using ReadyScript® cDNA Synthesis Mix to perform cDNA synthesis Tissue

Get tips on using First-Strand cDNA Synthesis Kit to perform cDNA synthesis Bacteria

Get tips on using SensiFAST™ cDNA Synthesis Kit to perform cDNA synthesis Bacteria

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. Multiplexing such a reaction amplifies the design challenges where one target requires 3 primers, which should be exclusively bound nowhere in the template DNA or to each other. Similarly, two targets require 6, three require 9, and so on. Each amplicon needs to be either a different size (for gels) or labeled with a different fluorescent tag that is spectrally distinct from the others in the reaction. Further complicating this, different targets in the reaction can compete with each other for resources and causes more challenges in the detection of amplicons. However, with proper primer designing, their validation, optimize quality and concentration of the enzyme and buffers certainly lead to a successful multiplex PCR reaction.