We do this routinely in a protocol that needs to recapture as much of the DNA as possible. To do this, we cut the band and place it in a 0.5 mL microcentrifuge tube with a hole punctured in its bottom by a 20-gauge needle. We place the tube with gel piece into a 1.7 mL microcentrifuge tube and spin for 1 min at max speed (13000 x g). This forces the gel through the tiny hole and shreds it into small pieces. To the shredded gel, we add 300 uL of the following "Soaking Solution":

2 mL 5M Ammonium Acetate

2 mL 1% SDS solution

4 uL 0.5 M EDTA

16 mL RNAse, DNAse free water

We incubate the gel pieces in this solution for 2 hrs at 70C with shaking. We briefly spin down the tube, transfer the supernatant (some gel chunk carryover is ok) into a Costar Spin-X Centrifuge Tube Filter, 0.22 um Cellulose Acetate column and spin at max speed for 1 min. To the flow-through we add 1 uL of 10 ug/uL glycogen, and then an equal volume (usually 300 uL) of isopropanol. We mix, then incubate overnight at -20C. Following incubation, we spin at 13,000 x g for 20 min, remove and discard the supernatant without disturbing the pellet, and add 200 uL of cold 80% ethanol. We again spin for 10 min at 13,000 x g, remove the supernatant without disturbing the pellet, and let air dry. The DNA pellet can be resuspended in water or the buffer of your choice. Typically, we get about 60-70% recovery by mass using this protocol which is high for gel-based methods.

Hi, have a look here. I think this will help you. http://www.protocol-online.org/cgi-bin/prot/view_cache.cgi?ID=1223

Cheers, Nadine

I put the cut band in water and vortexed, then centrifuged the gel down and took the supernatant. That was simple and worked well.

Cut the band into very small pieces, and put it into TE buffer for overnight. Then centrifuge and collect the supernatant.

If you did cut a band from a acrylamide gel, I would suggest transfer the band in 200 ul of sterile water in a 1.5 ml tube, incubate at 4 C overnight and the day after use 5 ul of that water as template to re-run the pcr.

Transfer the cut piece of gel in 200 microlitre MilliQ water (actually the quantity should be determined based on the intensity of your band) . Freeze thaw it twice. Centrifuge at 12000 x g for 10 minutes. Use the supernatant. good Luck.

Put the piece of gel on top of a Spin-X tube-filter (Corning), spin for 10 minutes and use the fluid to precipitate the DNA from. Spin-X centrifuge tube filters are used to filter (sterilize) small quantities.

Ethanol precipitation is the best way but make sure there is no residue. Tubes have to be completely dry with no alcohol traces...smell and add TE or Nuclease free water and shake it at 37 C for an hour then store at 4 C.

We do this routinely in a protocol that needs to recapture as much of the DNA as possible. To do this, we cut the band and place it in a 0.5 mL microcentrifuge tube with a hole punctured in its bottom by a 20-gauge needle. We place the tube with gel piece into a 1.7 mL microcentrifuge tube and spin for 1 min at max speed (13000 x g). This forces the gel through the tiny hole and shreds it into small pieces. To the shredded gel, we add 300 uL of the following "Soaking Solution":

2 mL 5M Ammonium Acetate

2 mL 1% SDS solution

4 uL 0.5 M EDTA

16 mL RNAse, DNAse free water

We incubate the gel pieces in this solution for 2 hrs at 70C with shaking. We briefly spin down the tube, transfer the supernatant (some gel chunk carryover is ok) into a Costar Spin-X Centrifuge Tube Filter, 0.22 um Cellulose Acetate column and spin at max speed for 1 min. To the flow-through we add 1 uL of 10 ug/uL glycogen, and then an equal volume (usually 300 uL) of isopropanol. We mix, then incubate overnight at -20C. Following incubation, we spin at 13,000 x g for 20 min, remove and discard the supernatant without disturbing the pellet, and add 200 uL of cold 80% ethanol. We again spin for 10 min at 13,000 x g, remove the supernatant without disturbing the pellet, and let air dry. The DNA pellet can be resuspended in water or the buffer of your choice. Typically, we get about 60-70% recovery by mass using this protocol which is high for gel-based methods.