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.
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.
Get tips on using CellTrace™ Calcein Red-Orange, AM to perform
Get tips on using 7-AAD (7-Aminoactinomycin D) to perform DNA quantification Human - BMDM
Get tips on using 7-AAD (7-Aminoactinomycin D) to perform DNA quantification Human - PC-3
Get tips on using REDExtract-N-Amp™ PCR ReadyMix™ to perform PCR Mouse
Get tips on using AmpliTaq Gold™ 360 DNA Polymerase to perform PCR Multiplex PCR - Bacterial DNA
Get tips on using Ion AmpliSeq™ Library Kit 2.0 to perform PCR Multiplex PCR - Mammalian DNA
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment