Some batches of light mineral oil can kill your PCR or RAPD reactions!

nishir at ohsu.edu nishir at ohsu.edu
Thu Jan 20 18:13:45 EST 1994


In article <pst-200194115408 at pst.pp.ksu.edu> pst at matt.ksu.ksu.edu (Paul St.
Amand) writes:
>We have recently found that a BRAND NEW gallon of light mineral oil
>was somehow preventing our PCRs and RAPDs from working.
>
>Has anyone else had this happen?
>
>We autoclaved the bad batch and some working oil and found that
>autoclaving did not restore the bad batch, nor did it cause the
>working batch to fail.
>
>Can anyone suggest why light mineral oil could prevent PCRs or RAPDs
>from working? Possible reasons: contamination, oxidation, light, or
>others?
>
>
>Paul St. Amand, pst at matt.ksu.ksu.edu, Voice: (913) 532-6168
>USDA & KSU Agronomy, 122 Throckmorton Hall, Manhattan KS 66506
>
We saw the same problem...variability with mineral oil from different vendors
and traced it to the stability of the mineral oil.  If you look carefully at
the labels, some are stabilized with vitamin E and some are not.  Apparently if
they are not stablilized, then they break down in the high temperatures of the
thermal cycler during PCR and create products that inhibit the PCR reaction. 
Our problems stopped as soon as we bought "vitamin E stabilized" mineral oil
(this is not on the label for the mineral oil, but part of the
ingredients/contaminants that are listed).

Rae Nishi
OHSU
Portland OR




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