5'end: protocol

Belhumeur Pierre belhumep at ERE.UMontreal.CA
Thu Apr 27 19:57:14 EST 1995


To all and more specifically to Pim and Preston,

I developped a 5' extension protocol during my years in the mammalian 
world an fortunately, it was reported in a paper of friends (hi Morrie and 
Frabncis) PNAS 263: 17987. I did primer extension few weeks ago using it (ON 
annealing at 25 C, in 50% formamide, as for RNase protection) with 100 
ug total RNA and, surprise, it worked fine!! One modification. Got rid of the
nucleotide (after labeling the primer) by 2 ammonium acetate precipitation
using glycogen as carrier. Not sure but everything went fine.

Hope it is helpful.

Pierre Belhumeur
Microbiology, U. of Montreal
belhumep at ere.umontreal.ca







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