PCR from blood dots on paper ?
jtNOjtSPAM at neurosearch.dk.invalid
Thu Feb 17 04:32:38 EST 2000
There should be enough template in a 1 cm blood dot for a large
number of reactions. I would dissolve the clots in a small
volume of TE buffer for long term storage at -20C.
The initial heating step in your PCR reaction should be
sufficient for lysing the cells. Run 30-35 cycles to get a
sufficient amount of product. Your cycle could look like this (a
standard PCR protocol):
96 C 5 min
96 C 30 s
55 C 30 s
72 C 1 min/kb of product
these 3 steps are repeated 30-35 times
72 C 5 min
If your main purpose is just to detect a band of a specific
size, conventional and cheap Taq should do the trick.
Dept. of Molecular Biology
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