PCR problems

vibeke vibekan at radium.uio.no
Thu Aug 16 03:39:07 EST 2001


Hello I am trying to amplify cDNA fragments (size 100bp-2000bp+). I am
doing this in 96 well plates. The problems are that I, when testing my
PCR reactions on agarose gels (1.5%), get high Mw smears on my gels. I
often get multiple bands as well. I am using universal primers and the
PCR conditions are: annealing 56*C, 1 min. and elongation 72*C, 3.5
min.

The PCR products are going to bee used in microarrays. Because of this
I need clean and high concentrated PCR products.

I would like to remove the high Mw smears on my gels, and maybe remove
some of the multiple bands.

Any useful hints would be greatly appreciated

Vibeke




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