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Resources

TRANSGENIC DNA PROTOCOL      

 

Transgenic DNA fragment purification protocol

1. Isolate DNA using maxiprep kit (do not pool minipreps)

2. Isolate tg fragment by restriction digest and run on a clean gel with clean buffer

3. Excise tg fragment from gel with clean razor blade

4. Purify tg fragment from gel using GENE CLEAN kit.

5. Resuspend DNA in tg buffer (7.5 mM TRIS pH 7.4, 0.2 mM EDTA, filtered through 0.2 um sterile filter) to a concentration of at least 50 ug/ul, with a volume of at least 50ul.

6. Spin Resuspended DNA through RPM spin filter (QBiogene, cat# 2080-800, extra catch tubes are 2080-801), make sure to prime filters with ~30ul tg buffer.

7. Run on a quantitative gel with mass ladder to check concentration and fragment purification, double check concentration on spec. You can freeze DNA sample at this point.