PCR of bugs in berries

Walter Hill walterh at U.WASHINGTON.EDU
Mon Jul 8 15:44:37 EST 1996


Hi

We're trying to develop a method to detect pathogenic microbes which
might be associated with various berries (e.g. straw, rasp, and
black) by using PCR.  We have found
that washes from berries have a strong inhibitory effect on PCR and
would like to know if anyone has any suggestions for removing
these inhibitors.  We wash the pellets extensively (3-4 washes)
before lysing the cells.  We have tried ethanol precipitating the DNA
but this leads to only a very slight improvement.  The procedure we
are using also includes Chelex so whatever is causing the inhibition
is not removed by it.  Anybody have any favorite columns or rapid DNA
prep methods that they would recommend?

TIA.  Regards, -Walt Hill


walterh at u.washington.edu (primary)      (secondary) whill at fdaem.ssw.dhhs.gov
Research Geneticist                         Seafood Products Research Center
U.S. Food and Drug Administration, Seattle District,  Bothell, WA 98041-3012
Affiliate Assoc. Prof. of Microbiology & Pathobiology    Univ. of Washington
VOICE:(206) 402-3176           "Cook it first"            (206) 483-4996:FAX




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