PCR with 32P-dCTP?

Bill Geese wjgeese at astro.ocis.temple.edu
Mon Nov 17 12:24:12 EST 1997


FOr a "typical" 50ul reaction (2mM MgCl2, 200uM dNTPs, 20pmol each primer, 
1ng plasmid template) when I use 32P-dCTp I add anywhere btw 10-50uCi 
(depending on the purpose) and cut the cold dCTP to 100uM. 


Bill

On 13 Nov 1997, gary vanzin wrote:

> Hello all;
> Has anyone worked out a protocol to PCR label DNA probes  using
> 32p-dCTP?  I tried it with our usual PCR protocol and it didn't work.
> I know that the radionuclotide (Amersham) comes in a solution containing 
> TEA,and this might prevent Taq from working correctly.  Any advice would
> be greatly appreciated.
> 
> ****************************
> Gary F. Vanzin
> University of Connecticut
> Department of Molecular and Cell Biology
> 75 North Eagleville Rd. U-125
> Storrs, CT 06268
> (860) 486-2101
> Fax: (860) 486-4331
> gfv94001 at uconnvm.uconn.edu
> ****************************
> 
> 
> 



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