Advances in multiplex PCR: balancing primer efficiencies and improving detection success.

ABSTRACT: 1. Multiplex PCR is a valuable tool in many biological studies but it is a multifaceted procedure that has to be planned and optimised thoroughly to achieve robust and meaningful results. In particular, primer concentrations have to be adjusted to assure an even amplification of all targeted DNA fragments. Until now, total DNA extracts were used for balancing primer efficiencies; however, the applicability for comparisons between taxa or different multiple-copy genes was limited owing to the unknown number of template molecules present per total DNA. 2. Based on a multiplex system developed to track trophic interactions in high Alpine arthropods, we demonstrate a fast and easy way of generating standardised DNA templates. These were then used to balance the amplification succ

SUBMITTER: Sint D 

PROVIDER: S-EPMC3573865 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature