PCR enhancer - (Aug/04/2005 )

It is recommended, if u go for PCR of larger fragment or having GC content more than 65% than better standarised your protocols with using BSA, DMSO,Glycerol or formamide.
Any body can tell me how these chemicals work or stabilise the things?

thanx in advance

I know for DMSO and formamide, they help reduce the secondary structures on the template (haripins for example) that would inhibit the reaction.

not too sure about BSA and glycerol, I thought these are used to preserve the Taq, but i maybe wrong.

Nick

I think formamide also decreases the annealing temperature? Can someone confirm? Does DMSO also decrease annealing temp?

Bovine serum albumin (BSA) usually help in amplification of ancient DNA or templates which contain PCR inhibitors such as melanin

Yes formamide and DMSO both decrease the Tm of the DNA. I have never had much luck with either and have found betaine to be much more useful.

Daniel

Automated DNA sequencing troubleshooting

At what concentration do you use formamide, betaine or DMSO?