POLYMERASE PRODUCTS |
RECOMMENDED FINAL PRIMER CONCENTRATION (EACH) |
FINAL PRIMER CONCENTRATION RANGE (EACH) |
---|---|---|
Q5® Polymerases |
500 nM |
200-1000 nM |
Phusion® Polymerases |
500 nM |
200-1000 nM |
OneTaq® Polymerases |
200 nM |
50-1000 nM |
Taq Polymerases |
200 nM |
50-1000 nM |
Hemo KlenTaq |
300 nM |
50-1000 nM |
LongAmp® Polymerases |
400 nM |
50-1000 nM |
We recommend a final primer concentration of 200 nM when using Taq-based polymerases, including OneTaq and EpiMark®.
Please refer to specific protocols on the product pages for details regarding range recommendations.
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Archaeal Family B polymerases, such as Q5 and Phusion, possess strong 3´-5´ exonuclease activity, which can digest nucleotides on the 3' end of primers and increase the likelihood of nonspecific amplification. Relative to Family A polymerases like Taq and OneTaq, a higher final primer concentration overcomes these effects and promotes specific product formation.
Generally, efficient PCR requires optimal ratios of magnesium, other ions, and primers to promote specificity by stabilizing the negative charges on the phosphate backbone. While the composition of Q5 and Phusion DNA Polymerases are proprietary, their buffers are optimized for using a final primer concentration of 500nM.