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PCR product on gel as smear - DNA isolated from paraffin blocks why smear? (May/12/2005 )

Hi,

I have been isolating DNA from paraffin embeded tissue using quiagen mini kit. I lysed few tissue sections in a standard buffer with proteinase K(0.4mg/ml) and keep it overnight . then i continue with quiagen kit protocol. My problem is out of the samples i have isolated after PCR for Viral DNA very few gives distint good bands while the rest are with dense smear. n out of these few good ones they stop amplifying after storing at 4deg C for one week . repeated freezing and thawing degrades DNA . I elute my DNA with AE buffer supplied with the KIT. Should i try with water . Pl help me resolve this two problem(smear as well as DNA storage).

thanks
remi

-p_reemi-

Hi Remi,

How big is your PCR product. remember that DNA from paraffin section is usually degraded and is hard to yield big product such as >500 bp.

-pcrman-