PCR on fresh tissue

casper vroemen Casper.Vroemen at MAC.MB.WAU.NL
Fri Jul 14 02:52:47 EST 1995


Dear Tsu-Hwie,

I have done lots of tissue PCR to detect GUS. Here's how:

- Place small piece of tissue, e.g. rosette leave, NO LARGER THEN 5x3 mm, in 0.5 ml eppendorf with 40 
ul of 0.25 M NaOH
- Boil 30 sec.
- Remove liquid and add 40 ul of 0.25 M HCl and 20 ul 0.5 M Tris-HCl 7.0/ 0.25% Nonidet-P40 (Sigma)
- boil 2 min.
- Remove all liquid and add 10 ul sterile water (this will be part of the reaction mixture)
- Perform PCR as usual

Casper Vroemen
Dept. of Mol. Biol.
Wageningen Agricultural Univ., the Netherlands



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