DNA Gel purification without Spin Column

[proatpcr]

Hello everyone

I have heard that many people can purify DNA from agarose gels without spin columns. Does anyone have a good method to do this?

thnx

[gandalfthegrey]

Hello again Pro,

We use electrolution apparatus to do this. You can purify DNA from agarose gel slices when even with small and large DNA fragments however many people use this with larger DNA from 5kb to 200Mb+.

[kmunson779]

I've done in-gel ligations by using low percentage (1-0.7%) low-melting temperature agarose, excising the band, melting it, and adding 1-2 uL to a 20 uL ligation reaction.

Depending on whether both insert and vector, or just one need to be gel purified, the finished ligation mixture may need to be melted before transformation.

There is also a freeze-squeeze method (that I've never personally done) that involves adding TE buffer, freezing, sometimes crushing, and either pipetting off the top or spinning through a homemade filtration apparatus to remove the gel particles. This protocol is usually used on acrylamide gels but could be done with agarose -- you can find details if you do a web search for "freeze-squeeze."