quiagen lambda preps
Large Hearted Boy
ed at bio02
Mon Feb 26 15:18:22 EST 1996
In an effort to increase my Lamda yields, I processed twice the
recommended amount of liquid lysate to be used per prep. I doubled up the
amount of L1 and L2 used, and after the PEG precipitation with L2, I
carried on as if I hadn't doubled the amount of lysate used. When I did
the Isopropanol precipitation, I immediately got a DNA clot that looked
suspiciously like genomic DNA. when I ran it out on a gel, although the
yield was great, there did indeed seem to be some genomic contamination.
Digests of the said DNA bore this out, as I got a tremendous smear, along
with the Lambda bands I was expecting. At first I wondered whether using
twice the amount of lysate could have caused the problem. I just wonder if
maybe the L1 has been sitting around too long and the DNAse I has gone bad.
Could I add some DNase I to the L1 I still have ?
thanks for any help you can provide me,
Dept. of Biology
University of Ottawa
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