What happens to 32P-dCTP when it goes KABLOOEY?

wolffie at u.washington.edu wolffie at u.washington.edu
Wed Mar 16 19:57:11 EST 1994


URGENT.  Answer required by Thurs. 3/17.

So I have all this stale (month-old) alpha-32P dCTP that I have to use  
because They consnabulated our nukes order.

What happens to an alpha-32P-labelled nucleotide triphosphate when the 32P   
decides it's time to disintegrate?  Does it blow completely to smithereens,  
leaving nothing to compete with the remaining 32P-dCTP in the labelling  
reaction, or does it leave some kind of mongrelized pyrophosphate with a  
nasty alpha-sulfate to bung up the works?

In other words, can I still get high specific activity in my probe by simply  
adding extra 32P-dCTP to the labelling reaction and/or letting it run  
longer, or does the Klenow enzyme utilize the spent 32P-dCTP, giving a  
colder probe?



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