Cell Transformation Group:Protocols/RNA and DNA - Revision history

Rhodri Harfoot at 00:02, 16 January 2009

2009-01-16T00:02:49Z

←Older revision		Revision as of 00:02, 16 January 2009
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-	==='''ETHANOL PRECIPITATION OF DNA'''===	+	==''ETHANOL PRECIPITATION OF DNA''==

	'''Materials:'''<BR>		'''Materials:'''<BR>

2009-01-16T00:01:17Z

←Older revision		Revision as of 00:01, 16 January 2009
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	{{Cell Transformation Group}}		{{Cell Transformation Group}}

-	===''ETHANOL PRECIPITATION OF DNA''===	+
		+	==='''ETHANOL PRECIPITATION OF DNA'''===

	'''Materials:'''<BR>		'''Materials:'''<BR>

2009-01-15T21:47:03Z

←Older revision		Revision as of 21:47, 15 January 2009
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	{{Cell Transformation Group}}		{{Cell Transformation Group}}
		+
		+	===''ETHANOL PRECIPITATION OF DNA''===
		+
		+	'''Materials:'''<BR>
		+	* 3 M sodium acetate pH 5.2 or 5 M ammonium acetate
		+	* DNA
		+	* 100% ethanol
		+
		+	1. Add 1/10 volume (of DNA sample) of 3 M sodium acetate, pH 5.2, or an equal volume of 5 M ammonium acetate. Mix well.
		+
		+	2. Add 2 to 2.5 volumes of cold 100% ethanol (calculated after salt addition) and mix well.
		+
		+	3. Place on ice or at -20°C for >20 minutes.
		+
		+	4. Spin at maximum speed in a microfuge 10-15 min.
		+
		+	5. Carefully decant supernatant.
		+
		+	6. Carefully add 1 ml 70% ethanol. Invert tube a couple of times to wash pellet.
		+
		+	7. Spin briefly if pellet too loose to tip off 70% ethanol straight away.
		+
		+	8. Tip off ethanol.
		+
		+	9. Spin again and pipette off last bit of 70% ethanol. Air dry tube up-side-down on tissue paper for a couple of minutes. Important not to over dry pellet or will be impossible to dissolve.
		+
		+	10. Resuspend pellet in the appropriate volume of TE or water by pipetting up and down. Can aid DNA resuspension by incubating at 65°C for 10-15min or at 37°C for longer.

2009-01-14T20:24:54Z

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