MSP question - (May/02/2006 )
Quick question. After poring through a number of papers which use MSP, i've noticed that a lot of people have published annealing temperatures at <55C and sometimes as low as 48-49. Wouldn't you expect a high frequency of mispriming at those temperatures? What would be considered "optimal" annealing temperatures to shoot for in MSP?
in short, the higher the better,
cancergeek, you are right in that mispriming will occur, after bisulfite treatment though it's difficult to pick primers with high Tm unless you extend the lengths of them to about 30mers. I would be a little suspicious with Tm's below 50C but if it works in the authors hands, it works!