Oligonucleotide Labeling with Polynucleotide Kinase

Dr. Peter Gegenheimer PGegen at UKans.nolospamare.edu
Fri Oct 1 20:47:24 EST 1999


On Mon, 27 Sep 1999 23:44:39, mlysik at hsc.vcu.edu wrote:

ð 	I am trying to label my oligonucleotides using gamma-32P-ATP and
ð polynucleotide kinase.  However, the labeling efficiency I get is low (often <1%) and I
ð get a high amount of incorporated label following purification.  I have used PNK from
ð various vendors and usually I add both excess oligo and ATP.

Another poster gave you the right protocol, but as a comment, you don't add
unlabeled ATP when you're shooting for high specific activity.

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