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[A convenient method of recovering DNA from agarose electrophoretic gels]
1College of Life Sciences, Beijing University, Beijing, 100871 P.R. China. wangl5@rpi.edu
Objective:
Construct a simple unit to efficiently recover DNA from agarose electrophoretic gels.
Methods:
With the use of 1.5ml microcentrifuge tube, 1 ml tip, nylon web membrane and dialysis membrane, the authors made a clever yet simple DNA recovering gadget, tested this gadget by using it to recover specific electrophoretic DNA bands of lambdaDNA/EcoR I+Hind III markers, and then examined the recovery ratio through agarose gel electrophoresis. A restriction enzyme's digestion product was taken as another sample to test the recovered product's quality by means of DNA ligation and subsequent agarose gel electrophoresis.
Results:
Under such electroelution conditions as with an electrophoretic buffer of 1xTBE, a voltage grade of 3V/cm and an elution time of 0.5h, the final recovery ratio of 564bp DNA was up to 80%, and that of 21227bp DNA was about 30%. Judged either by the agarose gel electrophoresis performance or by the DNA ligation result, the recovered DNA proved to be of excellent quality.
Conclusion:
It is an easy, quick, efficient and cheap method of recovering DNA from agarose electrophoretic gels. The recovering unit can be a permanent one, provided that the dialysis membrane in this unit is changed for recovering different DNA samples.