PCR product purification - (Apr/01/2004 )
I ususally purify three band, from the agarose gel containing the same PCR product, together to minimize the risk for diluting the sample.
seeing that your clock was ticking since april, not sure how much help my reply will be..
been using sod acetate- isoprop precipiation with no problems all the while.
good enough for cloning and gives good seq result when used as template..
let me know if u need the procedure
For cloning of PCR products, does phenol/chloroform extraction necessary? or can I just do a precipitation?
My two cents: With using a kit, to increase concentration while get good recovery, I usually use 25-30 ul buffer to elute DNA, after elution, use the same elutant to elute the same column once more.
Thanks all you guys for your numerous replies!!!
I have just repeated the PCR and obtained a better band... Next time I have a pool of choices here!!! But I am absolutely sure that this helped a lot of people. That makes this site great!!
Sounds like good old fashioned ethanol precipitation would do the trick.
Sometimes the the oldies are the best!