Download Plasmid DNA isolation (TENS prep) 1. Pour 1.5 ml overnight culture

Plasmid DNA isolation (TENS prep)
1.
2.
3.
4.
5.
6.
Pour 1.5 ml overnight culture into a microfuge tube
Spin at 10K for 1 min to pellet the cells
Pour out the supernatant, keep ~150 µl of media in the tube
Resuspend the cells by vortexing
Add 300 µl of TENS buffer
Invert the tube 3-4 times gently, the cells should lyse completely (the liquid turns
from turbid to clear)
7. Add 150 µl of 3M NaOAc (pH 5.6), invert 3-4 times gently, there should be white
precipitate forming
8. Spin at maximum speed for 5 min to pellet the white precipitate
9. Pipette the clear supernatant to a clean tube
10. Add 900 µl of 95% ETOH, invert tube to mix
11. Spin at maximum speed for 2 min to pellet DNA
12. Pour off the solution, then add 500 µl of 70% ETOH to wash the pellet by
vortexing
13. Spin again for 1 min.
14. Pour off 70% ETOH, dry the DNA pellet
15. Dissolve DNA in 50 µl 10mM Tris (pH 8) (Can be supplemented with RNase)
TENS buffer:
Tris-HCl
EDTA
NaOH
SDS
10 mM (pH 8.0)
1 mM
0.1 N
0.5% (w/v)