Miniprep/Qiagen kit - Revision history

Kyoung: /* Materials */

Kyoung — Fri, 06 Apr 2012 02:48:16 GMT

Materials

←Older revision		Revision as of 02:48, 6 April 2012
Line 5:		Line 5:

	For Qiagen buffer compositions, please see the [[Qiagen Buffers]] page.		For Qiagen buffer compositions, please see the [[Qiagen Buffers]] page.
		+	Note: Qiagen buffer also works for [http://www.epochlifescience.com/Product/SpinColumn/minispin.aspx Epoch Life Science's spin columns]which are sold in bulk at a much lower price. Yield and purity are quite close.

	==Protocol==		==Protocol==

Tk: /* Materials */

Tk — Sun, 31 Oct 2010 01:21:56 GMT

Materials

←Older revision		Revision as of 01:21, 31 October 2010
Line 4:		Line 4:
	For purifying plasmid DNA from ''Escherichia coli'' cells, the [http://www1.qiagen.com/Products/Plasmid/QIAprepMiniprepSystem/QIAprepSpinMiniprepKit.aspx Qiagen Spin Miniprep Kit] produces quite reliable results.		For purifying plasmid DNA from ''Escherichia coli'' cells, the [http://www1.qiagen.com/Products/Plasmid/QIAprepMiniprepSystem/QIAprepSpinMiniprepKit.aspx Qiagen Spin Miniprep Kit] produces quite reliable results.

-		+	For Qiagen buffer compositions, please see the [[Qiagen Buffers]] page.
-	~~Do not autoclave solutions containing isopropanol or MOPS; use sterile filtration if necessary.~~	+
-		+
-	~~Buffer P1~~	+
-		+
-	* 50 mM Tris-HCl pH 8.0	+
-	* 10 mM EDTA	+
-	* 100 μg/ml RNaseA	+
-		+
-	~~The buffer and RNaseA can also be ordered from~~ Qiagen ~~separately (catalog numbers 19051 and 19101).~~	+
-		+
-	~~Buffer P2~~	+
-	* 200 mM NaOH	+
-	* 1% SDS	+
-		+
-	~~Buffer P3 (not for spin columns, but for Qiatips, midi, maxi, giga kits)~~	+
-	* 3.0 M potassium acetate pH 5.5	+
-		+
-	~~Buffer N3~~	+
-	* 4.2 M Gu-HCl	+
-	* 0.9 M potassium acetate	+
-	* pH 4.8	+
-		+
-	~~Buffer PB~~	+
-	* 5 M Gu-HCl	+
-	* 30% ethanol	+
-	*(maybe add 10mM Tris-HCL PH 6.6, and that is better)	+
-		+
-	~~Buffer PE~~	+
-	* 10 mM Tris-HCl pH 7.5	+
-	* 80% ethanol	+
-		+
-	~~Buffer QBT equilibration~~ buffer	+
-	* 750 mM NaCl	+
-	* 50 mM MOPS pH 7.0	+
-	* 15% isopropanol	+
-	* 0.15% triton X-100	+
-		+
-	~~Buffer QC wash buffer~~	+
-	* 1.0M NaCl	+
-	* 50 mM MOPS pH 7.0	+
-	* 15% isopropanol	+
-		+
-	~~Buffer QF elution buffer~~	+
-	* 1.25M NaCl	+
-	* 50 mM Tris-HCl pH 8.5	+
-	* 15% isopropanol	+
-		+
-	~~Buffer QN~~	+
-	* 1.6M NaCl	+
-	* 50 mM MOPS pH 7.0	+
-	* 15% isopropanol	+
-		+
-		+
-	~~Buffer FWB2~~	+
-	* 1M potassium acetate, ~~pH 5.0~~	+
-		+
-		+
-	~~(Source:~~ [~~http://methodsandreagents.pbwiki.com/],~~ [~~[media:US6383393.pdf \| US Patent 6,383,393~~]])	+
-		+
-	~~The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9~~.3	+

	==Protocol==		==Protocol==

Vaishnavi Ananth: /* BioStream version */

Vaishnavi Ananth — Thu, 19 Nov 2009 10:00:57 GMT

BioStream version

←Older revision		Revision as of 10:00, 19 November 2009
Line 102:		Line 102:
	*Similarly, doing the elution in two steps (first a 30 μL elution and then a 20 μL dilution) can also slightly increase yields.		*Similarly, doing the elution in two steps (first a 30 μL elution and then a 20 μL dilution) can also slightly increase yields.

-	==~~BioStream~~ version==	+	==BioCoder version==
-	Following is the Miniprep/Qiagen kit 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 Miniprep/Qiagen kit 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====
	[[Miniprep/Qiagen kit protocol]]		[[Miniprep/Qiagen kit protocol]]

Vaishnavi Ananth at 09:39, 23 September 2009

Vaishnavi Ananth — Wed, 23 Sep 2009 09:39:15 GMT

←Older revision		Revision as of 09:39, 23 September 2009
Line 101:		Line 101:
	*Heating the elution buffer to 55°C prior to loading on the column can slightly increase yields.		*Heating the elution buffer to 55°C prior to loading on the column can slightly increase yields.
	*Similarly, doing the elution in two steps (first a 30 μL elution and then a 20 μL dilution) can also slightly increase yields.		*Similarly, doing the elution in two steps (first a 30 μL elution and then a 20 μL dilution) can also slightly increase yields.
		+
		+	==BioStream version==
		+	Following is the Miniprep/Qiagen kit 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====
		+	[[Miniprep/Qiagen kit protocol]]
		+	====Source Code====
		+	[[Miniprep/Qiagen kit protocol - source code]]

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

Torsten Waldminghaus at 17:07, 23 February 2009

Torsten Waldminghaus — Mon, 23 Feb 2009 17:07:15 GMT

←Older revision		Revision as of 17:07, 23 February 2009
Line 1:		Line 1:
		+	{{back to protocols}}
	==Materials==		==Materials==

Leo Tang: /* Materials */

Leo Tang — Sun, 22 Feb 2009 18:44:00 GMT

Materials

←Older revision		Revision as of 18:44, 22 February 2009
Line 29:		Line 29:
	* 5 M Gu-HCl		* 5 M Gu-HCl
	* 30% ethanol		* 30% ethanol
-	*(maybe 10mM Tris-HCL PH 6.6, and that is better)	+	*(maybe add 10mM Tris-HCL PH 6.6, and that is better)

	Buffer PE		Buffer PE

Leo Tang: /* Materials */

Leo Tang — Sun, 22 Feb 2009 18:43:23 GMT

Materials

←Older revision		Revision as of 18:43, 22 February 2009
Line 29:		Line 29:
	* 5 M Gu-HCl		* 5 M Gu-HCl
	* 30% ethanol		* 30% ethanol
		+	*(maybe 10mM Tris-HCL PH 6.6, and that is better)

	Buffer PE		Buffer PE

Tk at 14:54, 4 February 2009

Tk — Wed, 04 Feb 2009 14:54:47 GMT

←Older revision		Revision as of 14:54, 4 February 2009
Line 60:		Line 60:


-	(Source: [http://methodsandreagents.pbwiki.com/], ~~patent 6383392~~)	+	(Source: [http://methodsandreagents.pbwiki.com/], [[media:US6383393.pdf \| US Patent 6,383,393]])

	The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3		The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3

Tk: /* Materials */

Tk — Fri, 15 Aug 2008 21:01:22 GMT

Materials

←Older revision		Revision as of 21:01, 15 August 2008
Line 3:		Line 3:
	For purifying plasmid DNA from ''Escherichia coli'' cells, the [http://www1.qiagen.com/Products/Plasmid/QIAprepMiniprepSystem/QIAprepSpinMiniprepKit.aspx Qiagen Spin Miniprep Kit] produces quite reliable results.		For purifying plasmid DNA from ''Escherichia coli'' cells, the [http://www1.qiagen.com/Products/Plasmid/QIAprepMiniprepSystem/QIAprepSpinMiniprepKit.aspx Qiagen Spin Miniprep Kit] produces quite reliable results.

-	Buffer P1 ~~(in case you need to make it yourself) is:~~	+
		+	Do not autoclave solutions containing isopropanol or MOPS; use sterile filtration if necessary.
		+
		+	Buffer P1

	* 50 mM Tris-HCl pH 8.0		* 50 mM Tris-HCl pH 8.0
Line 47:		Line 50:
	* 15% isopropanol		* 15% isopropanol

		+	Buffer QN
		+	* 1.6M NaCl
		+	* 50 mM MOPS pH 7.0
		+	* 15% isopropanol
		+
		+
		+	Buffer FWB2
		+	* 1M potassium acetate, pH 5.0

Tk: /* Materials */

Tk — Fri, 15 Aug 2008 13:51:47 GMT

Materials

←Older revision		Revision as of 13:51, 15 August 2008
Line 14:		Line 14:
	* 200 mM NaOH		* 200 mM NaOH
	* 1% SDS		* 1% SDS
		+
		+	Buffer P3 (not for spin columns, but for Qiatips, midi, maxi, giga kits)
		+	* 3.0 M potassium acetate pH 5.5

	Buffer N3		Buffer N3
Line 27:		Line 30:
	* 10 mM Tris-HCl pH 7.5		* 10 mM Tris-HCl pH 7.5
	* 80% ethanol		* 80% ethanol
		+
		+	Buffer QBT equilibration buffer
		+	* 750 mM NaCl
		+	* 50 mM MOPS pH 7.0
		+	* 15% isopropanol
		+	* 0.15% triton X-100
		+
		+	Buffer QC wash buffer
		+	* 1.0M NaCl
		+	* 50 mM MOPS pH 7.0
		+	* 15% isopropanol
		+
		+	Buffer QF elution buffer
		+	* 1.25M NaCl
		+	* 50 mM Tris-HCl pH 8.5
		+	* 15% isopropanol
		+
		+

	(Source: [http://methodsandreagents.pbwiki.com/], patent 6383392)		(Source: [http://methodsandreagents.pbwiki.com/], patent 6383392)

	The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3		The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3
-

	==Protocol==		==Protocol==