Higher Yield Gel Extractions

Gel extractions can be a low yield affair, but overnight incubation over a spin column can increase yields. This protocol details a modified QIAquick Gel Extraction protocol.

QIAquick Gel Extraction Kit

  1. Cut the desired band out of an agarose gel using a scalpel.

  2. Weigh the slice in a snap cap tube and add 3 volumes of QG buffer per volume of gel, assuming the gel density is roughly 1µl/mg. This can be whatever gel dissolving buffer (with chaotropic salts and guanidine thiocyanate) you typically use.

  3. Add 1 volume of isopropanol to the tube and vortex.

  4. Place a QIAquick spin column in a collection tube. Add the sample to the column and incubate over column overnight at room temp.

  5. Centrifuge spin column for 1 minute at 13,000 RPM.

  6. Discard flow through. Repeat step 5 if the sample volume was greater than 800 µl.

  7. Add 750 µl of buffer PE to column and centrifuge for 1 minute at 13,000 RPM. Discard flow through.

  8. Centrifuge for an additional 1 minute to remove all wash buffer.

  9. Place column in collection tube and add 50 µl EB buffer or water directly to the silica portion of the column. Incubate up to 4 minutes then centrifuge the column for 1 minute to elute DNA.

Analyze DNA using a standard Nanodrop/general spectrophotometer protocol for 260/280 absorbance ratios. Store all samples in a -20 oC freezer.

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Lily Ben-Aviover 1 year ago

Awesome, thanks so much for posting Amos! I have always dreaded gel extractions, and look forward to trying this out. I will also add that warming the EB buffer or water to 56˚C before elution has consistently improved my yield!

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