Can protocols optimized for Phusion® DNA Polymerase directly be applied to Phusion® Hot Start DNA Polymerase?
In many cases yes. However, with Phusion® Hot Start DNA Polymerase, use primers with Tm 60 ºC or higher. Typically the length of such primers is 20 nt or more. Calculate the Tm values with the nearest-neighbor method (see a link to a calculator below). In addition, use of detergent free reaction buffers is not recommended with Phusion® Hot Start DNA Polymerase. Tm calculator for PCR.
Related Questions
- Do DyNAzyme™ I, II and EXT DNA Polymerases, and Phusion® Polymerase add the non-template dependent 3-A overhang?
- Can protocols optimized for Phusion® DNA Polymerase directly be applied to Phusion® Hot Start DNA Polymerase?
- Does Phusion® Hot Start DNA Polymerase need a separate activation step in the PCR protocol?
