As I am currently pursuing a new junior faculty position, the Yale URL and email may change in the near future. The information provided in these pages is copyrighted and is intended for individual use only. Click here to get the Biotechniques paper in PDF format. Although equimolar primer mixtures did not usually provide optimal amplification of all loci, this test allowed the observation that too high and too low primer amounts may need to be avoided .Too high primer concentrations may inhibit the multiplex reaction whereas too low amounts may not be sufficient.
Numerical values indicate the concentration of each primer in the final reaction.
Mixture A includes 14 primers and mixture B includes 10. Within limits, increasing primer concentration may improve the outcome of the PCR reaction, and should be considered as a way to optimize PCR reactions. This is exemplified in Fig. 14 below, in which increasing primer and template DNA concentrations, both improve the results. PCR with various concentrations of genomic DNA template and various concentrations of a degenerate primer.