Purification of small (100bp) PCR products
Posted 11 March 2005 - 04:22 AM
Does anybody have a good purification protocol for small (~100bp) PCR products?
Posted 11 March 2005 - 09:21 AM
Posted 11 March 2005 - 01:48 PM
And their prices are cheaper too.
Posted 12 March 2005 - 08:34 PM
You can run your PCR on a agarose gel, cut the band of interest out, place into an eppy and add 100uL of TE. Centrifuge to ensure the slice of gel is in TE, or you can now mash the gel up with a pipette tip. Then place the tube into a 60C heat block for about 15 minutes. Then spin the tube at maximum on a table top centrifuge for 20 minutes. Aspirate the supernatant which now contains only the DNA of interest.
Edited by methylnick, 12 March 2005 - 08:35 PM.
Posted 13 March 2005 - 03:23 AM
well methylnick, i have to do a pcr tomorrow, i will try your protocol!!!
Posted 13 March 2005 - 09:19 AM
Hi Nick, will there any problem for such extracted DNA in downstream applications such as ligation, sequencing...
Thanks you guys for sharing.
Posted 13 March 2005 - 08:35 PM
good luck with it!
I have managed to ligate the product into a TA vector, sequence, do nested PCR and restriction digest it.
I think as long as the amount of supernatant used is minimal it should be fine.
Good luck with it!