PCR from human tissues

nswema05 at angis.su.OZ.AU nswema05 at angis.su.OZ.AU
Wed Jan 24 16:43:08 EST 1996


I have performed PCR on human peripheral blood mononuclear cells and
skin samples. We handled all blood samples and tissues in a biosafety
cabinet in a category 2 cell biology laboratory. Surgical gowns were
worn over laboratory coats, overshoes were worn and two pairs of
gloves were worn while working with samples. The outer pair of gloves
was removed whenever the hands were taken out of the biosafety cabinet
and another pair put on before the hands reentered the cabinet.
Tubes were vigorously sprayed with 70% alcohol or 1% Virkon before
being opened; although this may not destroy contaminating DNA, it may
contribute to overall cleanliness in the sense of diluting or washing
away contaminants. Clean sterilised equipment, tubes and reagents were
used at all steps. We tended to use new items whenever possible.
Plugged tips are important for use with pipettes.

PCR reactions mixes were prepared in a separate room, preferably again
using a biosafety cabinet with similar precautions being taken. These
were brought into our cell biology laboratory to be added to the
samples. The PCR thermal cycling was done in a machine outside the
cell biology laboratory and PCR products were handled in a separate
molecular biology laboratory. This minimised the risk of PCR product
contamination of samples.




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