DNA Preparation, Plasmid DNA - TELT Method


Jeff Lawrence

1. Grow cells overnight in LB at 37° with shaking.

2. Microfuge cells for 30 sec. Remove supernatant by aspiration.

3. Resuspend cells in 100 µL TELT Buffer:
	50 mM	Tris, pH 8.0	(5 mL	1 M Tris, pH 8.0)
	62.5 mM	EDTA, pH 8.0	(12.5 mL 500 mM EDTA, pH 8.0)
	2.5 M	LiCl		(31.5 mL 8 M LiCl)
	4 %	Triton X-100	(4 mL	Triton X-100)
				(53 mL	H2O)
4. Add 100 µL 25:24:1 Phenol/Chloroform/Isoamyl alcohol.

5. Vortex vigorously for 15 sec.

6. Centrifuge for 1 min at room temperature.

7. Transfer supernatant to a new microcentrifuge tube.

8. Add 200 µL 95% Ethanol. Mix well and incubate at room temperature for 2 min.

9. Microfuge for 5 min at 4°.

10. Discard supernatant. Add 1 µL 70% Ethanol, swirl, and discard.

11. Air dry.

12. Resuspend in 20 µL TE.
Last Update: Thursday, 19-Jun-2014 11:39:50 PDT
This page has been viewed 89 times.
Eric Kofoid eckofoid at ucdavis.edu