PCR Problems

Rudi test at czzorlnm.azn.nl
Thu Aug 26 09:55:00 EST 1999


How did you check your reagents I ask myself? The fact that you are getting
a product in your control lane when it contains no DNA-template is the only
way to check for contamination. Neither gel-electroforesis nor
spectrofotometrie will show the low amounts of DNA that are sufficient for
contamination (think pico or femto molar range.).
To get rid of contamination in water-controls replace all reagents with new
ones, make new primer-dilutions and clean your pipets inside and out. Also,
is your taq-polymerase clean? I have seen a message in this group stating
that the taq-enzyme always contains trace amounts of (bacterial) DNA. Are
you trying to amplify a bacterial gene?

Rudi van de Wetering


"Deborah Sims" heeft geschreven in bericht ...
>Hi
>
>I am an honours student having problems with my PCR reactions.  I keep
>getting amplification in my control lane which is supposed to have water
>in instead of DNA.  I have checked for contamination but all the
>reagents appear to be clean! Is it possible that the primers are somehow
>being amplified?  Any comments or advice would be greatly appreciated.
>
>Thank You
>Debbie Sims





More information about the Methods mailing list