Contamination woes...HELP!

Robert DeLisio QuintMark at earthlink.net
Mon Jun 8 22:30:56 EST 1998


Decomtamination of pipets with UV is not effective.

The UV light source must be perpendicular to the surface to be
decomtaminated in order to inactivate amplifiable DNA. Pipets are 3-D and
have rounded surfaces that are not exposed to sufficient UV to completely
decontaminate. I've found that wiping down pipets ( or any other surface )
with 10% bleach is the most effective way to get rid of contaminating DNA.

Bleach should be used with caution, however. Should ANY bleach get into your
reaction mixture it will kill it completely.

Regards,
Bob DeLisio

helen.turner at nri.org wrote in message <6l0nh4$5cl$1 at nnrp1.dejanews.com>...
|In article <6kum38$qs0$1 at news1.bu.edu>,
|  rkoedood at bu.edu (Marieke R. Koedood Zhao) wrote:
|>
|> I got that in my early PCR days. First PCR beautiful, next one all were
|> positive. What aved me after that was not using the same pipets for
setting
|> up the reaction and analyzing the product. Also set up my reactions
|> on a different bench (or even lab) as the one where I analyze the PCR
product.
|>
|> Marieke
|>
|
|I quite agree that keeping separate sets of pipettes for PCR set-up
|and analysis is a good idea.  If this is not possible, then do at least
|try to ensure that:
|
|a) set-up and analysis are carried out as far from
|each other as possible (separate rooms is best !) and
|
|b) use tips with filters in, to prevent sucking up DNa
|into the pipette or expelling contaminants already present into new PCR
|reactions.
|
|In the meantime, bleach your benchtops and decontaminate your pipettes with
|UV.
|
|Helen.
|
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