Mouse tissue lysis for genotyping - Revision history

Vaishnavi Ananth at 11:33, 22 March 2010

2010-03-22T11:33:40Z

←Older revision		Revision as of 11:33, 22 March 2010
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	* 4°C short term, -20°C long term		* 4°C short term, -20°C long term
		+
		+
		+	==BioCoder version==
		+	Following is the Mouse tissue lysis for genotyping 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====
		+	[[Mouse tissue lysis for genotyping protocol]]
		+	====Source Code====
		+	[[Mouse tissue lysis for genotyping protocol - source code]]

	== see also ==		== see also ==

Jakob Suckale: /* see also */

2008-01-16T15:37:52Z

Jakob Suckale: categories

2008-01-16T15:21:38Z

Jakob Suckale: backlink

2008-01-16T15:14:41Z

backlink

←Older revision		Revision as of 15:14, 16 January 2008
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		+	{{back to protocols}}
		+
	This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.		This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.

Jakob Suckale: links

2008-01-16T15:14:12Z

links

←Older revision		Revision as of 15:14, 16 January 2008
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	* 4°C short term, -20°C long term		* 4°C short term, -20°C long term

-	== ~~links~~ ==	+	== see also ==

	* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]		* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]
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	* [[Designing primers]] - how to design primers		* [[Designing primers]] - how to design primers
	* [[PCR techniques]] - overview page		* [[PCR techniques]] - overview page
		+
		+	== external links ==
		+	* [[Image:3stars.png]] [http://cancer.ucsd.edu/tgm/TailGenomicDNAPrep.pdf UCSD protocol genomic DNA from mouse tails]
		+	* [[Image:2stars.png]] [http://www.bio.com/protocolstools/protocol.jhtml?id=p1689 DNA extraction from tail or tissue] - at BioProtocol from unknown contributor
		+	* [http://www.protocol-online.org/prot/Molecular_Biology/DNA/DNA_Extraction___Purification/DNA_Extraction_from_Cell_and_Tissue/Tail_DNA_Extraction/ Protocol Online - tail DNA extraction protocols]
		+

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:In vivo]]		[[Category:In vivo]]
	[[Category:Mouse]]		[[Category:Mouse]]

Reshma P. Shetty at 21:25, 8 May 2007

2007-05-08T21:25:38Z

←Older revision		Revision as of 21:25, 8 May 2007
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	* [[Designing primers]] - how to design primers		* [[Designing primers]] - how to design primers
	* [[PCR techniques]] - overview page		* [[PCR techniques]] - overview page
		+
		+	[[Category:Protocol]]
		+	[[Category:In vivo]]
		+	[[Category:Mouse]]

Jakob Suckale: temp 56 to 50

2007-05-02T14:48:56Z

temp 56 to 50

←Older revision		Revision as of 14:48, 2 May 2007
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-	* incubate for 3h to o/n at ~~56°C~~ at 300 rpm	+	* incubate for 3h to o/n at 50°C at 300 rpm
	:(proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C)		:(proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C)

Jakob Suckale: material, prot K stability

2007-05-02T14:48:14Z

material, prot K stability

←Older revision		Revision as of 14:48, 2 May 2007
Line 1:		Line 1:
	This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.		This is a quick protocol for mouse tail and tissue lysis with proteinase K. It is commonly used to prepare templates for genotyping. Other protocols included detergents in the lysis buffer, but we found this simple protocol to work well with less hands-on time.

-	==== tissue lysis to release DNA ====	+	== material ==
		+	* proteinase K
		+	* Taq 10x buffer
		+	* tabletop shaker/incubator
		+
		+	== procedure ==
		+	=== tissue lysis to release DNA ===
		+
		+	per tail or tissue chunk add (tissue degradation is sped up if tissue/tail is cut into smaller bits):
		+	* 100 µl Taq DNA polymerase 10x Reaction buffer with MgCl2 (e.g. Promega #M1883) diluted 1:10 in ddH2O
		+	* +2 µl proteinase K (20 mg/ml)
		+

-	* per tail or tissue chunk (tissue degradation is sped up if tissue/tail is cut into smaller bits):
-	** 100 µl Taq DNA polymerase 10x Reaction buffer with MgCl2 (e.g. Promega #M1883) diluted 1:10 in ddH2O
-	** +2 µl proteinase K (20 mg/ml)
	* incubate for 3h to o/n at 56°C at 300 rpm		* incubate for 3h to o/n at 56°C at 300 rpm
		+	:(proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C)

-	==== proteinase K inactivation ====	+	=== proteinase K inactivation ===

	* 95°C for 10-20 min at 300 rpm		* 95°C for 10-20 min at 300 rpm
	* centrifuge at 13000 rpm for 3 min and transfer supernatant (80 µl) into new eppi		* centrifuge at 13000 rpm for 3 min and transfer supernatant (80 µl) into new eppi

-	==== storage before PCR ====	+	=== storage before PCR ===

	* 4°C short term, -20°C long term		* 4°C short term, -20°C long term

-	=== links ===	+	== links ==

	* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]		* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]

Jakob Suckale: /* proteinase K inactivation */

2007-04-18T17:25:51Z

proteinase K inactivation

←Older revision		Revision as of 17:25, 18 April 2007
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	==== proteinase K inactivation ====		==== proteinase K inactivation ====

-	* 95°C for 20 min at 300 rpm	+	* 95°C for 10-20 min at 300 rpm
	* centrifuge at 13000 rpm for 3 min and transfer supernatant (80 µl) into new eppi		* centrifuge at 13000 rpm for 3 min and transfer supernatant (80 µl) into new eppi

Jakob Suckale: links

2007-04-18T13:35:53Z

links

←Older revision		Revision as of 13:35, 18 April 2007
Line 16:		Line 16:

	* 4°C short term, -20°C long term		* 4°C short term, -20°C long term
		+
		+	=== links ===
		+
		+	* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]
		+	* [[PCR]] - how to do a PCR
		+	* [[Designing primers]] - how to design primers
		+	* [[PCR techniques]] - overview page

←Older revision		Revision as of 15:37, 16 January 2008
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	== see also ==		== see also ==

		+	* [[DNA extraction from tissue]]
	* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]		* [http://en.wikipedia.org/wiki/Genotyping Genotyping in Wikipedia]
	* [[PCR]] - how to do a PCR		* [[PCR]] - how to do a PCR

←Older revision		Revision as of 15:21, 16 January 2008
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	[[Category:In vivo]]		[[Category:In vivo]]
	[[Category:Mouse]]		[[Category:Mouse]]
		+	[[Category:DNA]]