DNA extraction from gel - (Apr/18/2007 )
How do I extract PCR product from gel after electrophoresis? Are there many or standard method in doing so?
The easiest way for you to extract your PCR product from gel is using a commercially available kit. The protocol will be ready together with the kit.
search for low melting temperature gel extraction method.
electroelution may be a point. Then you have commercial kits or beta-agarase which digest low meting agarose gels.
1. Gel extraction (easiest is from a kit.)
2. PCR purification (from a kit). Run a small sample out on a gel to ensure it's the product you want. After that you purify the remaineder of the PCR product. Nice and easy.
i prefer the PCR purification just because I lose less product that way.
Some kits (like the Promega Wizard SV) can do both gel extraction and PCR cleanup, so it's pretty good value. We re-use the columns too (after washing them of course!) so it's really very cheap.
use EASY TRAP ver 2 for gel extraction
its depend upon you further experiment.
you can use silica based DNA purification or passing through column.
Both kits available in the market.
In the case of colum you get pure eluted DNA and in the case of silica there may be some purity which interfare with your further enzymatic reaction.
thats why you decide before purchasing any kits from market.
The columns are also silica based. And YES, they do release some sort of residue. Try running water through the column and measure before + after with a spectro. It's enough to throw off the yield calculation (if you have a very low yield), so you should keep some 'eluted water' as a blank.
for cloning, i get far better results not to do gel purif.
I gel purify a digested template, and after the PCR i do a column or silica matrix beads.