Reasons for PCR failure....?

Sailesh Surapureddi Sailesh.Surapureddi at
Fri Aug 2 11:25:43 EST 1996


        Here some of the reasons that I picked up for PCR, which might be
helpful to you.

1. Clean template DNA (preferably Qiagen purified) @ low conc in sterile water
   or TE.

2. DEPC free water,, after long struggling.. I have found that Autoclaved
   Q water to be the best.

3.  Re calibrate your pipettes. (if the calibration is screwed up, you
might be
    adding excess of primers etc etc and there goes your PCR to dogs). In
    case, I guess this is the problem, since you have written everything
    about the pipettes

4. Powder free gloves (if you happen to wear any, most of the  times the powder
   from the gloves tend to inhibit reactions).

5. Avoiding the formation of bubbles while pipetting (a common mistake, usually

Last but not the least TONS of GOOD LUCK (..just kidding)


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