Team:Hawaii/Protocols/gel extraction

Gel Extraction

 * Using Qiagen MiniElute Gel Extraction Kit

Protocol

 * 1) Cut band(s) from gel.
 * 2) Determine weight of gel.
 * If weight of gel >400mg, run as 2+ extractions in different tubes.
 * 1) Add Buffer QG.
 * For gels <2% agarose, add 3 gel volumes of QG.
 * For gels >2% agarose, add 6 gel volumes of QG.
 * 1) Incubate at 50C, shaking every 2-3 minutes, until gel is dissolved.
 * 2) Check the color of the solution is yellow. If the solution is orange, add 3M sodium acetate until solution is yellow (indicates neutral pH).
 * 3) Add 1 gel volume of isopropanol and mix by inverting.
 * 4) Transfer solution to elution spin column (purple).
 * 5) Centrifuge 1 min. at room temperature at 13,000 rpm.
 * 6) Discard flow through.
 * 7) Add 500 &mu;l Buffer QG.
 * 8) Centrifuge 1 min. at room temperature at 13,000 rpm.
 * 9) Discard flow through.
 * 10) Add 750 &mu;l Buffer PE and incubate 2 min.
 * 11) Centrifuge 1 min. at room temperature at 13,000 rpm.
 * 12) Discard flow through.
 * 13) Transfer spin column to fresh eppendorf tube.
 * 14) Add 10 &mu;l Buffer EB and incubate 1 min.
 * Add buffer directly onto filter in the spin column.
 * 1) Centrifuge 1 min. at room temperature at 13,000 rpm.
 * 2) Collect flow through and store at -20C.