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 June 19 2014
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Eric Kofoid
eckofoid at ucdavis.edu