Size selective DNA precipitation - Revision history

Sander B. Frank at 22:13, 6 January 2012

2012-01-06T22:13:53Z

←Older revision		Revision as of 22:13, 6 January 2012
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	*5% -- <700bp		*5% -- <700bp
	*4% -- <1kb <br>		*4% -- <1kb <br>
-	~~<br>~~	+
	Notes:		Notes:
	**Yield decreases considerably when <10% PEG is used		**Yield decreases considerably when <10% PEG is used
	**Always use 10mM final concentration of MgCl<sub>2</sub>		**Always use 10mM final concentration of MgCl<sub>2</sub>
-
-
	==Troubleshooting==		==Troubleshooting==

2012-01-06T22:10:33Z

←Older revision		Revision as of 22:10, 6 January 2012
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	*4% -- <1kb <br>		*4% -- <1kb <br>
	<br>		<br>
-	Notes: ~~<br>~~	+	Notes:
-	:Yield decreases considerably when <10% PEG is used	+	**Yield decreases considerably when <10% PEG is used
-	:Always use 10mM final concentration of MgCl<sub>2</sub>	+	**Always use 10mM final concentration of MgCl<sub>2</sub>

2012-01-06T22:09:33Z

←Older revision		Revision as of 22:09, 6 January 2012
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	==Other PEG Concentrations and Approximate Size Exclusion==		==Other PEG Concentrations and Approximate Size Exclusion==

-	*% PEG Fragments Excluded	+	% PEG -- Fragments Excluded <br>
	*10% -- <300bp		*10% -- <300bp
	*6% -- <500bp		*6% -- <500bp
	*5% -- <700bp		*5% -- <700bp
-	*4% -- <1kb	+	*4% -- <1kb <br>
-		+	<br>
-	*Note1: Yield decreases considerably when <10% PEG is used	+	Notes: <br>
-	*Note2: Always use 10mM final concentration of MgCl<sub>2</sub>	+	:Yield decreases considerably when <10% PEG is used
		+	:Always use 10mM final concentration of MgCl<sub>2</sub>

2012-01-06T22:02:53Z

←Older revision		Revision as of 22:02, 6 January 2012
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	*% PEG Fragments Excluded		*% PEG Fragments Excluded
-	*10% -- <300bp	+	*10% -- <300bp
	*6% -- <500bp		*6% -- <500bp
	*5% -- <700bp		*5% -- <700bp
	*4% -- <1kb		*4% -- <1kb

-	*~~Note~~: Yield decreases considerably when <10% PEG is used	+	*Note1: Yield decreases considerably when <10% PEG is used
		+	*Note2: Always use 10mM final concentration of MgCl<sub>2</sub>

2012-01-06T22:00:56Z

←Older revision		Revision as of 22:00, 6 January 2012
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	==Critical steps==		==Critical steps==
	*Before centrifugation mark the tubes in order to know where the pellet will be expected afterwards, as the pellet will be (nearly) invisible		*Before centrifugation mark the tubes in order to know where the pellet will be expected afterwards, as the pellet will be (nearly) invisible
		+
		+	==Other PEG Concentrations and Approximate Size Exclusion==
		+
		+	*% PEG Fragments Excluded
		+	*10% -- <300bp
		+	*6% -- <500bp
		+	*5% -- <700bp
		+	*4% -- <1kb
		+
		+	*Note: Yield decreases considerably when <10% PEG is used
		+

	==Troubleshooting==		==Troubleshooting==

2009-11-19T09:49:46Z

BioStream version

←Older revision		Revision as of 09:49, 19 November 2009
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	Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.		Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.

-	==~~BioStream~~ version==	+	==BioCoder version==
-	Following is the Size selective DNA precipitation 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 Size selective DNA precipitation 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====
	[[Size selective DNA precipitation protocol]]		[[Size selective DNA precipitation protocol]]

2009-10-20T10:16:56Z

←Older revision		Revision as of 10:16, 20 October 2009
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	Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.		Tag this page with categories to allow easier indexing and searching. See [[Categories]] for information on existing categories.
		+
		+	==BioStream version==
		+	Following is the Size selective DNA precipitation 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====
		+	[[Size selective DNA precipitation protocol]]
		+	====Source Code====
		+	[[Size selective DNA precipitation protocol - source code]]

	[[Category:Protocol]] [[Category:DNA]] [[Category:In_vitro]]		[[Category:Protocol]] [[Category:DNA]] [[Category:In_vitro]]

2009-02-19T11:54:06Z

←Older revision

Revision as of 11:54, 19 February 2009

2008-08-27T23:08:00Z

Notes

←Older revision		Revision as of 23:08, 27 August 2008
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	==Notes==		==Notes==
-	*This protocol is being ~~mention~~ in the manual for the Gateway recombinational cloning system based on published methods.<cite>Website Paithankar Lis</cite>	+	*This protocol is being mentioned in the manual for the Gateway recombinational cloning system based on published methods.<cite>Website Paithankar Lis</cite>
	*Adjusting the PEG concentration can shift the threshold of the size of the precipitated DNA. The higher the final PEG concentration, the smaller the fragments that will be removed (which will stay in the supernantant).		*Adjusting the PEG concentration can shift the threshold of the size of the precipitated DNA. The higher the final PEG concentration, the smaller the fragments that will be removed (which will stay in the supernantant).
	*If the sample volume is different, simply adjust the other volumes accordingly to end up with the same ratio.		*If the sample volume is different, simply adjust the other volumes accordingly to end up with the same ratio.

2008-08-27T23:07:16Z

Reagents

←Older revision		Revision as of 23:07, 27 August 2008
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	===Reagents===		===Reagents===
-	*DNA to be separated	+	*DNA to be separated (e.g. PCR reaction mixture)
	*30% (w/v) PEG 8000/30 mM MgCl<sub>2</sub> (concentration of PEG 8000 can be varied to shift the size of the percipitated DNA. The concentration used here will remove DNA fragments with less than 300bp)		*30% (w/v) PEG 8000/30 mM MgCl<sub>2</sub> (concentration of PEG 8000 can be varied to shift the size of the percipitated DNA. The concentration used here will remove DNA fragments with less than 300bp)
	*TE Buffer, pH 8.0 (10 mM TRIS-HCl, 1 mM EDTA, pH 8.0)		*TE Buffer, pH 8.0 (10 mM TRIS-HCl, 1 mM EDTA, pH 8.0)