PCR from blood dots on paper ?

T.Hiltunen hiltunen at csc.fi
Wed Feb 16 01:57:01 EST 2000


Hello!

We have blood dots (two 1-cm dots from each subject) on 
paper and aim to do PCR with them. I ask for your help in 
deciding the best way to proceed, since even the Red Book
does not tell the right procedure. Two options come into my mind:

1) We (this means the technician ;) excise a small (1mm x 1mm) 
   piece of paper for each PCR reaction and start the reaction 
   with prolonged denaturation. Is the 9-min 95°C used with 
   AmpliTaq Gold sufficient ?

2) We incubate the whole blood dot in 1xPCR buffer and use an 
   aliquot in the PCRs. Does this require boiling or something 
   else? How is the DNA stored? Any tricks in PCR parameters?

Any help is appreciated.

With best regards,

Timo Hiltunen
MD, PhD
Dept of Medicine
University of Helsinki
Finland




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