18 (May 18, 2011 GDS)

(Difference between revisions)

Revision as of 20:29, 9 September 2011

TENS How-To

4.5 TE

250λ 10% SDS

250λ 2M NaOH

Mini-Preps (How-To)

1. Spin 1.5 mL of overnight culture for 30 secs. in micro-centrifuge.

2. Aspirate off all but 200uL of supernatant and resuspend the pellet by vortexing.

3. Add 300uL of TENS and mix by inversion. The solution should become viscous.

4. Add 150uL of sodium acetate and vortex. A fine white precipitate should form.

5. Centrifuge for 2.5 minutes.

6. Transfer the supernatant to clean tube and add 2 volumes (1mL) of room temp ETOH.

7. Mix and pellet DNA by centrifugation for 2-5 min.

8. Wash pellet with 70% ethanol and allow pellet to dry.

9. Resuspend the pellet in 30uL TE.

10. Digest 5-10uL as usual.

Minipreps (2 each) for:

pNICE SA-15λ TE

J024430 GE-15λ TE

Trans002-5, AB-Resp 30λ

Trans003-W1 SD, SA-

@@ Line 1: / Line 1: @@
-Minipreps (How-To)
-Parth--made fresh TENS
+TENS How-To
 :::4.5 TE
 :::250&lambda; 10% SDS
@@ Line 7: / Line 6: @@
-::Spin 1.5 mL (2 tubes) 30"
+Mini-Preps (How-To)
-::Pour off sol'n + resuspend
-::Add: 300&lambda; TENS mix
-:::150&lambda; NaOAc mix
-::Spin 2.5'
+. Spin 1.5 mL of overnight culture for 30 secs. in micro-centrifuge.
-::Sup to new tube
-::Add 1 mL ETOH - mix well
+. Aspirate off all but 200uL of supernatant and resuspend the pellet by vortexing.
-::Spin 3' ________
-::Wash 70% ETOH
+. Add 300uL of TENS and mix by inversion. The solution should become viscous.
+. Add 150uL of sodium acetate and vortex. A fine white precipitate should form.
+. Centrifuge for 2.5 minutes.
+. Transfer the supernatant to clean tube and add 2 volumes (1mL) of room temp ETOH.
+. Mix and pellet DNA by centrifugation for 2-5 min.
+. Wash pellet with 70% ethanol and allow pellet to dry.
+. Resuspend the pellet in 30uL TE.
+. Digest 5-10uL as usual.