G-tailing ss cDNA.

Mr D J Plows lsren at csv.warwick.ac.uk
Fri Feb 4 14:19:19 EST 1994


Dear all, 

	I'm having trouble with adding a G tail to my single stranded
cDNA. I suspect that the buffer that comes with the enzyme is not the 
one I want. Accordin to the bible of molecular biology (Maniatis, sp!),
tailing of ss cDNA requires either Mg2+ or Mn2+ depending upon what 
nucleotide is to be added. However, the buffer that comes with the 
terminal transferase has Co2+ (I think it's 2+) in it. According to the 
bible this buffer is used when tailing ds DNA. SO, the question is, does
anyone have a recipe for terminal transferase buffer that contains either
Mg2+ or Mn2+. BTW, the cDNA works fine in PCR's when using internal primers.
Any suggestions would be of help. 

	Thanks in advance.

	DAve.




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