How to 32P label a PCR probe?

CK Wen cwen at aux.btny.purdue.edu
Fri Jan 12 17:04:21 EST 1996


bina at unixg.ubc.ca (Jim) wrote:
>Hi, Does anyone have a simple receipe for labelling a DNA probe with 32P-ATP in 
>a PCR reaction?
>
>Thanks,
>Jim
>
This is my reaction:

Plasmid DNA, which contains an insert     2-5ng     1ul
10X PRC buffer                                      1ul
10XdNTP(dATP, dTTP and dGTP) 50uM                   1ul
dCTP-P32                                            5ul
Primer 1, 50-100pg/ul                               1ul
Primer 2                                            1ul
Taq                                                 1ul

Then go to PCR.

The probe made is very hot, for more information, please check
http://expert.cc.purdue.edu/~ckwen/  
under" in Molecular Biology" where all the hybridization were performed using PCR generated probe.

best wishes,

Chi-Kuang.




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