FAQ: What PCR reagents do you recommend for DNA amplification in genome editing (CRIPSR/Cas9,TALEN,ZFN) mismatch detection assays?

We recommend Q5® High-Fidelity DNA Polymerase with Q5® Reaction Buffer or Q5 Hot Start High Fidelity 2X Master Mix because of Q5 DNA Polymerases’ high fidelity and robustness.
The EnGen Mutation Detection Kit provides reagents for detection of on-target genome editing events.  The kit comes with optimized enzymes (including PCR reagents), protocols and includes a control.

Notes: 

  1. T7 Endonuclease I is a structure-selective enzyme. It acts on a variety of DNA substrates with different specific activities. It is important to control the amount of enzyme and the reaction time used for cleavage of a particular substrate.
  2. Temperatures above 42°C cause an increase in nonspecific nuclease activity and should be avoided.
  3. pUC(AT) is derived from pUC19 with a modification of the polylinker between the EcoRI site and the PstI site.