Rapid DNA Preparation for Restriction Analysis

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℃ for 30 sec.,then cool to 4℃

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)