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UV Nicking of DNA in Agarose Gels for Enhanced Transfer and Detection of Megabase Size DNAs

sportation to the GS Gene Linker chamber, place the tray inside the GS Gene Linker chamber, and close the door.

3. Irradiate the gel with 60 mJoules of energy (nic program). The gel can now be photographed, but further exposure to UV light must be minimized (<10 seconds). The gel can be destained prior to photography if desired.

4. Soak the gel in 0.4 N NaOH, 1.5 M NaCl for 15 minutes. Transfer the DNA for at least 16 hours to Zeta- Probe GT membrane according to standard procedures, using 0.4 N NaOH, 1.5 M NaCl as the transfer solvent.

This protocol is for use with the GS Gene Linker UV chamber, with gels approximately 6 mm thick. If thicker gels are used, the staining period should be prolonged. DNA that is not stained with EtBr will not be nicked by UV irradiation, and will not transfer efficiently.


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