SNGE,NIMH,NIH
http://intramural.nimh.nih.gov/lcmr/snge/Protocols/Miniprep2.html
| SOLUTIONS:
- Green and blue solutions from Clone Checker kit (cat. no. 11666-013).
PROCEDURE:
- Place 3-6µl of an overnight culture in an Eppendorf tube (or spread part of a bacterial colony on the bottom of tube with a toothpick, which then can be used to create new colony on another plate). Add 8µl of green solution. Vortex
- Heat at 100°C for 30 sec., then cool to 4°C
- Add 2µl 10x restriction buffer, enzymes and water to the cell preparation to 20µl total.
- Incubate at the appropriate restriction enzyme temperature for 30 min.
- Add 2µl stop dye with RNase (blue solution), mix and run on an agarose gel.
- (Note: if the gel is stained briefly after the run in ethidium bromide (0.5µg/ml), any genomic DNA smear may appear less prominent than if the gel were run in buffer containing ethidium bromide)
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