PCR without mineral oil

[CH...@gr.forth.imbb.nefelh]

If you have the money you can use "Ampliwax" I've used it once, and since
it solidifies at room temperature, there are no contamination problems. I
am also told you can reduce primer artefacts by loading primers above the
solidified wax plug. The folks in my lab here have a neat trick to remove
parafin oil using "Parafilm". Take your finished PCR reaction and remove
as much as you can from beneath the oil. Spot on to a piece of parafilm
and roll the liquid back and forth a few times. Parafin oil sticks to the
film leaving the aqueous material as a separate blob. Just pipette it
back up again.
Cheers,

Rod

[Bob Ludwig lab]

re mineral oil

I add 75 ul cholorform to a PCR reaction and sometimes
a little dilute bromphenol blue loading dye. The chloroform
mixes with the oil and makes it denser than the aqueous.
The blue makes the aqueous phase very easy to see. With
a little sucrose in the dye, it will pipette directly into the
well of an agarose gel. It can also be re-PCR'd
--b0b kuhn
------------------------------------------------
Robert Kuhn, PhD
Sinsheimer Laboratories
University of California
Santa Cruz, CA 95064 USA

ph: 408-459-2728
fax: -3139
email: rk...@orchid.ucsc.edu
================================================