Cut out gel slice with the desired DNA fragment and weigh gel.
Add 1:1 volume of binding buffer to gel slice. (Eg. 100μL of binding buffer for every 100mg of agarose gel.)
Incubate gel solution at 50°C-60°C for 10 minutes. Inverting tube helps melting process.
(Optional: only use if DNA fragment is <500bp or >10kb long) If DNA fragment is <500bp, add a 1:2 volume of 100% isopropanol to the solubilized gel solution. Mix thoroughly. If DNA fragment is >10kb, add a 1:2 volume of water to the solubilized gel solution. Mix thoroughly.
Binding DNA
Transfer (up to 800μL) gel solution to spin column.
Centrifuge for 30-60 seconds. Discard flow-through and place spin column back into same collection tube.
Washing column of residue
Add 700μL of wash buffer and centrifuge for 1 minute.
Discard flow-through and the centrifuge empty column for 1 minute.
Place the column into a fresh 1.5mL microcentrifuge tube.
We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!
We would also like to thank and acknowledge: Our Advisors
Marc Facciotti
Ilias Tagkopoulos Technical Guidance
David Larsen
Andrew Yao Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China) cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)
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