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Revision as of 20:02, 4 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
-	:Minipreps (2 each) for:
-	:pNICE SA-15λ TE
-	:J024430 GE-15λ TE
-	:Trans002-5, AB-Resp 30&lambda
-	:Trans003-W1 SD, SA-
-	::Spin 1.5 mL (2 tubes) 30"	+	Mini-Preps (How-To)
-	::Pour off sol'n + resuspend	+
-	::Add: 300λ TENS mix	+
-	::::        150λ NaOAc mix	+	1. Spin 1.5 mL of overnight culture for 30 secs. in micro-centrifuge.
-	::Spin 2.5'	+
-	::Sup to new tube	+	2. Aspirate off all but 200uL of supernatant and resuspend the pellet by vortexing.
-	::Add 1 mL ETOH - mix well	+
-	::Spin 3' ________	+	3. Add 300uL of TENS and mix by inversion. The solution should become viscous.
-	::Wash 70% ETOH	+
		+	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.