FAQ: I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Q5® High-Fidelity DNA Polymerase?

  • Use more template. Sample concentration may be too low. 
  • Template DNA may be damaged. Use carefully purified template. 
  • Optimize enzyme concentration by testing a titration of enzyme in the reaction (0.25-2 units/50μl reactions) 
  • Increase number of cycles. 
  • Lengthen extension time to 40s/kb.