Fuzzy PCR product band

Jens Tornoe jens at tornoe.net
Fri Mar 3 17:37:40 EST 2000


I have made the same observation after building dsDNA from two ~150 base
randomized oligos with complemetary 3' ends using Taq. The resulting band
appeared to be approximately the right size, but even if I didn't add Taq, a
band could still be seen on the gel. It was fainter, but had the same size.
All bands were very fuzzy. Column purification helped a bit.

We really couldn't explain what we saw, but since the synthesized dsDNA
could be cloned and the sequences were correct, I hurried on with the work
trying my best to forget that.

Jens Tornøe
NsGene, Denmark



AlPiEs <alpies at aol.com> wrote in message
news:20000303093810.10719.00003200 at ng-da1.aol.com...
> We are constructing a phage-based peptide library and have amplified a 123
b
> oligonucleotide containing a 90 b randomized sequence using PCR.   When we
run
> the product in a 1% agarose gel we get a "fuzzy" band, in the 200-300 bp
range.
>  Is this normal? I thought that it might be due to the heterogeneity of
the
> product. Thanks.
> Alan Escher






More information about the Methods mailing list