Team:Paris Saclay/mini maxi

From 2013.igem.org

Revision as of 20:06, 22 September 2013 by Damip (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Mini et maxi preparation

1. Centrifuge during 1 minute the cellular culture in tubes of 1.5 or 2.0 mL at maximal speed

2. Add 100µL of solution n°1 (50 mM glucose, 10 mM EDTA and 25 mM Tris pH 8.0) and hang up the residue (culot)

3. Add 200µL of solution 2, buffer solution of lyse (0.2 mM NaOH et 1% SDS), mix the final solution until it is translucent

4. Add 150µL of solution 3, precipitation with potassium acetate 3.3M, incubate in ice during 10min

5. Centrifugation at maximal speed during 10 min

6. Take off the supernatant liquid and transfer it in a new tube

7. Add 500µL of phenol chloroforme

8. Centrifuge at maximal speed during 8 min, take off the aqueous phase

9. Precipitation with ethanol