Maxiprep of plasmid DNA from E. coli - Revision history

Vaishnavi Ananth at 09:36, 19 November 2009

2009-11-19T09:36:56Z

←Older revision		Revision as of 09:36, 19 November 2009
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-	==~~BioStream~~ version==	+	==BioCoder version==
-	Following is the Maxiprep of plasmid DNA from E.coli protocol in ~~BioStream~~, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in ~~BioStream~~ (see Source code). More information about ~~BioStream~~ can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]	+	Following is the Maxiprep of plasmid DNA from E.coli protocol in BioCoder, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioCoder (see Source code). More information about BioCoder can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
	====Text Output====		====Text Output====
	[[Maxiprep of plasmid DNA from E.coli protocol]]		[[Maxiprep of plasmid DNA from E.coli protocol]]

Vaishnavi Ananth at 06:27, 23 September 2009

2009-09-23T06:27:13Z

←Older revision		Revision as of 06:27, 23 September 2009
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	You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.		You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.
		+
		+
		+	==BioStream version==
		+	Following is the Maxiprep of plasmid DNA from E.coli protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
		+	====Text Output====
		+	[[Maxiprep of plasmid DNA from E.coli protocol]]
		+	====Source Code====
		+	[[Maxiprep of plasmid DNA from E.coli protocol - source code]]

	[[Category:Protocol]]		[[Category:Protocol]]

Torsten Waldminghaus at 17:05, 23 February 2009

2009-02-23T17:05:41Z

←Older revision		Revision as of 17:05, 23 February 2009
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		+	{{back to protocols}}
	This protocol produces very clean, concentrated DNA, usually on the order of several mg/mL. It is labor intensive. Expect to spend most of the day doing it, but I like to store my plasmids in this form.		This protocol produces very clean, concentrated DNA, usually on the order of several mg/mL. It is labor intensive. Expect to spend most of the day doing it, but I like to store my plasmids in this form.

-	===Ingredients===	+	==Ingredients==
	Ingredients are per culture; make enough for one extra culture to allow for pipetting error).		Ingredients are per culture; make enough for one extra culture to allow for pipetting error).

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	* 125μL 3M sodium acetate		* 125μL 3M sodium acetate

-	===Directions===	+	==Directions==

	#Grow a single colony of E. coli overnight in 50mL LB broth + selective markers at 37°C.		#Grow a single colony of E. coli overnight in 50mL LB broth + selective markers at 37°C.
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	#Centrifuge at 13,600g/12,000rpm, 4°C, for 15 minutes. Discard the supernatant. Wash pellet with ~100μL 70% ethanol. Resuspend in 100-200μL TE buffer.		#Centrifuge at 13,600g/12,000rpm, 4°C, for 15 minutes. Discard the supernatant. Wash pellet with ~100μL 70% ethanol. Resuspend in 100-200μL TE buffer.

-	===Notes===	+	==Notes==

	You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.		You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.

Madhadron at 11:43, 25 September 2007

2007-09-25T11:43:39Z

←Older revision		Revision as of 11:43, 25 September 2007
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	[[Category:DNA]]		[[Category:DNA]]
	[[Category:In vitro]]		[[Category:In vitro]]
		+	[[Category:Escherichia coli]]

Madhadron: Maxiprep moved to Maxiprep of plasmid DNA from E. coli: More precise name

2007-09-25T11:41:29Z

Maxiprep moved to Maxiprep of plasmid DNA from E. coli: More precise name

←Older revision

Revision as of 11:41, 25 September 2007

Reshma P. Shetty at 19:38, 27 April 2007

2007-04-27T19:38:12Z

←Older revision		Revision as of 19:38, 27 April 2007
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	You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.		You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.

-	[[Category:Protocol]] [[Category:~~Escherichia coli~~]] [[Category:~~DNA~~]]	+	[[Category:Protocol]]
		+	[[Category:DNA]]
		+	[[Category:In vitro]]

Reshma P. Shetty at 19:31, 27 April 2007

2007-04-27T19:31:40Z

←Older revision		Revision as of 19:31, 27 April 2007
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	You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.		You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.
		+
		+	[[Category:Protocol]] [[Category:Escherichia coli]] [[Category:DNA]]

Madhadron at 22:42, 24 March 2007

2007-03-24T22:42:46Z

←Older revision		Revision as of 22:42, 24 March 2007
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	#Add 750μL straight ethanol and 125μL 3M sodium acetate. Put at -80°C for 30 minutes or -20°C overnight.		#Add 750μL straight ethanol and 125μL 3M sodium acetate. Put at -80°C for 30 minutes or -20°C overnight.
	#Centrifuge at 13,600g/12,000rpm, 4°C, for 15 minutes. Discard the supernatant. Wash pellet with ~100μL 70% ethanol. Resuspend in 100-200μL TE buffer.		#Centrifuge at 13,600g/12,000rpm, 4°C, for 15 minutes. Discard the supernatant. Wash pellet with ~100μL 70% ethanol. Resuspend in 100-200μL TE buffer.
		+
		+	===Notes===
		+
		+	You may want to read [http://www.madsci.org/posts/archives/2000-07/965055056.Mb.r.html this discussion] of what LiCl and sodium acetate are doing in this protocol.

Madhadron: /* Ingredients */

2007-03-17T23:07:32Z

Ingredients

←Older revision		Revision as of 23:07, 17 March 2007
Line 8:		Line 8:
	* 111μL 20mg/mL lysozyme		* 111μL 20mg/mL lysozyme
	* 2mL worth of components for solution 2: 200μL 10% SDS, 100μL 4M NaOH, 1.7 mL autoclaved water		* 2mL worth of components for solution 2: 200μL 10% SDS, 100μL 4M NaOH, 1.7 mL autoclaved water
-	* 1.5mL Solution 3: 3M K<sup>+</sup>, 5M acetate (3M potassium-acetate, 2M acetic acid --- glacial is 17M)	+	* 1.5mL Solution 3: 3M K<sup>+</sup>, 5M acetate (3M potassium-acetate, 2M acetic acid -- glacial is 17M)
	* 3.7mL isopropanol		* 3.7mL isopropanol
	* 1mL TE buffer		* 1mL TE buffer

Madhadron: /* Ingredients */

2007-03-17T23:07:24Z

Ingredients

←Older revision		Revision as of 23:07, 17 March 2007
Line 8:		Line 8:
	* 111μL 20mg/mL lysozyme		* 111μL 20mg/mL lysozyme
	* 2mL worth of components for solution 2: 200μL 10% SDS, 100μL 4M NaOH, 1.7 mL autoclaved water		* 2mL worth of components for solution 2: 200μL 10% SDS, 100μL 4M NaOH, 1.7 mL autoclaved water
-	* 1.5mL Solution 3: 3M K<sup>+</sup>, 5M acetate (3M potassium-acetate, 2M ~~glacial~~ acetic acid)	+	* 1.5mL Solution 3: 3M K<sup>+</sup>, 5M acetate (3M potassium-acetate, 2M acetic acid --- glacial is 17M)
	* 3.7mL isopropanol		* 3.7mL isopropanol
	* 1mL TE buffer		* 1mL TE buffer