PCR product purification - (Apr/01/2004 )

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I ususally purify three band, from the agarose gel containing the same PCR product, together to minimize the risk for diluting the sample.

/Nina

-Nina-

seeing that your clock was ticking since april, not sure how much help my reply will be..

anywayz,

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..

good luck!

let me know if u need the procedure

-julianne-

QUOTE

for PCR product purification used for cloning, phenol/chloroform's classical method is ok. I think kits are unnecessary.

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.

-postdoc-

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!!

-thegradstudent-

Sounds like good old fashioned ethanol precipitation would do the trick.
Sometimes the the oldies are the best!

-tuckern-

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