DNA Transfer - Plaque/Colony Lift

Jeff Lawrence

1. Wet circular Nitrocellulose filter in TE and place on chilled agar plate. Orient the filter on the plate by puncturing the filter asymmetrically with a needle.

2. After 4 min carefully lift the filter from the plate and transfer it to Whatman 3MM paper saturated with:

	0.5 M	NaOH	10 mL	5 N NaOH
	1.0 M	NaCl	20 mL	5 M NaCl
			70 mL	ddH20

3. After 4 min, transfer the filter to Whatman 3MM paper saturated with 1 M Tris, pH 7.5.

4. After 4 min, transfer filter to Whatman 3MM paper saturated with:

	0.5 M 	Tris, pH 7.5	50 mL	1 M Tris, pH 7.5
	1.5 M	NaCl		30 mL	5 M NaCl
				20 mL	ddH₂O

5. Transfer to a piece of dry Whatman 3MM paper and air dry.

6. Bake at 80° for 90 min.

Last Update: Thursday June 19 2014

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Eric Kofoid eckofoid at ucdavis.edu