How to decide the concentration of Polymerase for fidelity assays?

08 January 2018 1 1K Report

So, at lower concentration of polymerase 20nM , the polymerase shows only 40% incorporation . The conc of DNA 50nM, dNTP 100 micromolar. Can I go for higher concentration of polymerase to show fidelity? OR what else can be changed so that I can keep the lower concentration of polymerase . I would like to know the range of Polymerase and dNTPs which are permissible concentartions ranges.

Oded Babai

Some times to much is not so good. Try to teduce DNA or primers. Playing with magnesium can help. You have to try many combination to get the ideal preformances

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