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PCR primers: hairpin and dimers? - what can I do to avoid this? (Dec/15/2005 )

hello All smile.gif
I have designed primers for PCR but after checking I understood that they can both form hairpin structures and self dimers.....i cant change the primer sequence so I will order the primers but can I do anything to avoid hairpin formation or dimer formation later on in my PCR. thanx a lot!

-Kathy-

If faulty primer design is unavoidable, hot-starting PCR (manual or use hot-start enzymes), using PCR additives such as DMSO may help.

-mario2004-

please i want to know if i have this problem than my PCR is going to fail or maybe not.....sorry new to this feild....

-Kathy-

QUOTE (Kathy @ Dec 19 2005, 03:15 PM)
please i want to know if i have this problem than my PCR is going to fail or maybe not.....sorry new to this feild....


as suggested above if u cannot change the sequence of the primer (which would be the easiest thing to do) it would be necessary to use a hot start Taq and keep the annealing temp fairly high - you could also try using touch down PCR where u start froma high annealing temp and decrease by 0.5oC per cycle for a number of cycles and hoep that u may get a small amount of product forming.

Regards

-scientist555-

its always hard to predict if your PCR is going to work or not, having sub-optimal primers does not help.

Along with other suggestions from the rest - you could also try using really small amounts of your primer during the PCR... but i agree re-designing the primers would be easiest, adn might even be cheaper at the end of it all!

-janbrisbane-