5' end labeling

Vadim Kraynov vkraynov at pop.service.ohio-state.edu
Sun Jun 8 17:22:33 EST 1997

In article <aisoai-0806970441220001 at ppp112.yokohama2.dtinet.or.jp>, aisoai at agc.co.jp (ISOAI Atsushi) wrote:
>"A. Kumar" <akumar at blue.weeg.uiowa.edu> wrote:
>>I am having trouble with 5' end labeling of a 28mer oligo. I am using a
>>molar ratio of 32P to substrate DNA of 3:1, and the specific activity of
>>the 32P-ATP is 6000 Ci/mmol. I was getting good labeling about a year
>>and all of a sudden it stopeed working. If anyone has any suggestions or
>>ideas, I would really appreciate it.
>Why not you discard all reagents including enzymes and
>oligonucleotides? I recommend you should purchase fresh
>reagents. Bye.

Exactly.  Buy new kinase and try again. Also new 32P ATP.  I do this on a 
dayly basis and it ALWAYS works.

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