HOT-Start-PCR: How do I make wax?

raymond Henderson samhaoir at HOTMAIL.COM
Wed Jun 17 08:21:05 EST 1998


Jochen wrote:

Currently, I try to enhance my RT-PCR method by doing Hot-Start-PCR.
But the manual method (pipetting a key reagent) is not very
practical, especially when you want to run many reactions at the same
time.
Therefore I want to use wax as an separator of my PCR-Components.
But as you may know, commercial waxes as AmpliWax (R) or others are
extremely
expensive ($80 and more for 200 Pieces). Also, anti-taq-antibodies
are much to expensive, too. So I have following QUESTION:
Has anybody made these wax-pellets manually? What materials, beside
paraffine, do I use?


I've found 1 part paraffin wax to 3 (or4) parts mineral oil to be very 
effective DNA/Taq barrier for Hot-Start. This mix melts at 60-70 oC and 
solidifies at 35-40 oC. I've carry out 96 PCR reactions simultaneously 
on several occasions using this method with no mis-priming apparent.

Raymond Henderson
Genetic Vectors, Inc.

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