I run an electrophoresis in agarose gel and my expected bands are extremely week, is even hard to see them. It looks I need to increase my PCR efficiency. I have read about a Hot start setp, which means pre-heat all my mix with my DNA at 95º (holding step it looks) and after that, add my taq polymerase. Did anyone try that before?

Apart of that and increasing MgCl2 concentration, is there anything else I can do? maybe increase my DNA volumen?

  • $\begingroup$ More cycles seems to me the trivial way to go. $\endgroup$ – cagliari2005 Jul 13 '18 at 5:04
  • $\begingroup$ I have several questions of your prep. How many cycles are you doing? What are your pcr mix components (concentrations/amounts)? What is your current setup procedure? What are your cycle temps? What primers are you using and what DNA are you using? $\endgroup$ – tswei Jul 13 '18 at 13:33

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