horton at cis.umn.edu
Fri Mar 31 22:13:49 EST 1995
G Shafer (geshafer at aol.com) wrote:
: In article <3lcibn$bv4 at news.acns.nwu.edu>, jkh141 at nwu.edu wrote:
: > Does anyone sell cheap wax beads for hot-starting PCR? I
: > tried Histo embedding wax beads, but I had to chop them up with
: > a razor blade and wasn't keen on this potential source of
: > contamination.
: > Jody K. Hirsh
: > Northwestern University, Chicago, IL. USA
: > jkh141 at nwu.edu
: I haven't tried it, but you might check out the following:
: R.M.Horton, B.L.Hoppe, B.M.Tronconi(1994)BioTechniques vol 16, 42-43
: "Ampligrease:'Hot Start" PCR using petroleum jelly"
Try it, you'll like it!
The bottom mix is covered with vaseline (aka "AmpliGrease") by dispensing
with a syringe, melting, and cooling. Then the top mix is added, and the
whole mess is covered with oil. If you use enough oil (about 3 times the
volume of the grease), the oil/grease mix will not solidify after cooling.
A protocol for hot start in 10 ul volumes is included. The whole thing
works better if you heat up the sample block before you put the samples
in (so they start *real* hot).
Bob Horton (Ph.D.!) /\ "Crash programs fail because of the theory that
U. of Minnesota, CBS || with nine women pregnant you get a baby a month"
1479 Gortner Ave. /||\ -Werner von Braun. Disclaimer:"Bob who?"
St. Paul, MN 55108 ^^ horton at molbio.cbs.umn.edu/(612) 624-3790
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