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Methods for primer reconstitution - (May/03/2006 )

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Well, if you are doing reverse transcript reactions, you DO want nuclease free water. And I disagree with the "just water" approach. Pure water is acidic (or will become so with the CO2 dissolving into it). Low pH is bad for DNA. You want a buffer, at pH 7.5 to 8.0. EDTA helps if you have DNAse present, since it chelates Mg++ from solution. There is little reason not to use TE, since primers are used in such small molar amounts and are typically kept at high concentration.


Ok guys,

This thing is making me go nuts! blink.gif

Can I use the TE buffer(Tris-Hcl pH 7.4,EDTApH8.0)?I use this buffer to reconstitute my DNA.Would it be safe to use it for PCR purposes as well becoz normally they recommend to keep stuff of DNA prep.and PCR separate due to contamination issues.What say I save one unused bottle of TE from the kit and use it exclusively for reconstitution of primers?


OK relax...all right, here's the gist of all these posts and various theories, that I can see

there is more than one way to do this, and people have varying opinions on the best way. I personally agree with phage434, but that does not mean it is the only way that will work. I suspect you could resuspend your primers in the TE that you have. segregating your buffers is a good thing, especially if you are new to PCR...and use very very careful technique when you get into/out of those little bottles and you should be OK

or, you can use water. it's really up to you...just make sure it's very clean

good luck


I am gonna use nuclease free water (promega).That settles it!


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