DNA-binding protein cloning

Wei Shao", ADDRESS: IN%"w_shao at uvmvax.uvm.edu W_SHAO at UVMVAX.BITNET
Tue Dec 8 16:28:00 EST 1992


Hi there:
I am trying to clone a DNA-binding protein from Lambda-gt11 cDNA library.
I have a question about the concentrations of labelled oligo probe used
in probing step. According to Sambrook et al "Molecular Cloning", the best
result can be obtained with 25 ng probe/ml (specific activity is 2x10-8
cpm/ug). That is, 5x10-6cpm/ml. Most other protocols call for 1x10-6
cpm/ml. According to your experienct, what kind concentration is better?
Thanks in advance.
W Shao
Dept of Microbiology and Molecular Genetics
Univ of Vermont
w_shao at uvmvx.uvm.edu
USA



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