dsOligos annealing buffer
Posted 04 March 2005 - 08:23 PM
This is Sung. I am right now working on annealing two complementary single stranded oligos about 60mers. From what I have found, in order to anneal the oligos, I need to use annealing buffer. However, instead of the annealing buffer, does anybody know if I can just use PCR buffer from Roche?
Posted 07 March 2005 - 01:32 AM
i did not try this one. So if you do so, please tell us your results.
For information, here is my recipe :
2µg of each oligo
Buffer STE (10x) :
tris HCl pH 7,5 100mM
NaCl : 500mM
EDTA pH 8 : 10mM
heat your oligos 5' at 95° and let the temperature slowely decrease to room temp.
Posted 07 March 2005 - 03:53 AM
2 ug Forward oligo
2 ug Reverse oligo
Up to 50 ul with TE
Heat @ 90ºC for 3 min
Incubate @37ºC for 1 h