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.
More information about the Methods
mailing list