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From 2011.igem.org

Revision as of 21:24, 3 September 2011 (view source) Sallen (Talk | contribs) ← Older edit		Latest revision as of 15:08, 24 September 2011 (view source) Sallen (Talk | contribs)
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-	Parth--made fresh TENS	+	TENS How-To:
-	::4.5 TE	+	:::4.5 TE
-	::250λ 10% SDS	+	:::250λ 10% SDS
-	::250λ 2M NaOH	+	:::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.
		+
		+
		+	<gallery widths=910px heights=800px>
		+	File:Page0005.jpg
		+	</gallery>

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Latest revision as of 15:08, 24 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.