In vitro transcription with T7 RNA polymerase - Revision history

Jakob Suckale: /* External links */

Jakob Suckale — Sat, 20 Jun 2009 15:07:55 GMT

External links

←Older revision		Revision as of 15:07, 20 June 2009
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	* [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)]		* [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)]
	* [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion]		* [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion]
-	* [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia]	+	* [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia], [http://www.fermentas.com/catalog/modifyingenzymes/rnapolymerases.htm RNA polymerases - Fermentas]
-		+

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:RNA]]		[[Category:RNA]]
	[[Category:In vitro]]		[[Category:In vitro]]

Jakob Suckale at 15:02, 20 June 2009

Jakob Suckale — Sat, 20 Jun 2009 15:02:23 GMT

←Older revision		Revision as of 15:02, 20 June 2009
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	In vitro transcribed RNAs like those from T7 or similar viral promoters like T3 and SP6 are important components for many molecular biology experiments. They can be used to generate (antisense) RNA probes for blot hybridisation and nuclease protection assays that are more sensitive than randomly primed DNA probes. Modified nucleotides containing isotopes like <sup>32</sup>P or detectable epitopes like DIG can be integrated into the RNA via T7 transcription. Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments.		In vitro transcribed RNAs like those from T7 or similar viral promoters like T3 and SP6 are important components for many molecular biology experiments. They can be used to generate (antisense) RNA probes for blot hybridisation and nuclease protection assays that are more sensitive than randomly primed DNA probes. Modified nucleotides containing isotopes like <sup>32</sup>P or detectable epitopes like DIG can be integrated into the RNA via T7 transcription. Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments.

-	~~== Lab-specific protocols ==~~	+	The [[Sauer lab]] has an excellent, detailed protocol: [[Sauer:In vitro transcription with T7 RNA polymerase]].
-	* [[Sauer:In vitro transcription with T7 RNA polymerase]]	+
-	* [[Knight:In vitro transcription]]	+	For a detailed description of PCR-based attachment of T7 promoters see [[Making RNA probes with T7 transcription]].
		+
		+	== See also ==
		+	* [[Knight:In vitro transcription\|Knight:In vitro transcription with E. coli RNA polymerase]]

	== Related OWW pages ==		== Related OWW pages ==

Jakob Suckale: back to protocols

Jakob Suckale — Sat, 20 Jun 2009 14:50:55 GMT

back to protocols

←Older revision		Revision as of 14:50, 20 June 2009
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		+	{{back to protocols}}
		+
	'''In vitro T7 transcription''' is the synthesis of RNAs using a T7 promoter and purified enzyme. It is the standard method of making up to several mg of RNAs longer than about 20nts with relatively high quality as compared to solid phase synthesis.		'''In vitro T7 transcription''' is the synthesis of RNAs using a T7 promoter and purified enzyme. It is the standard method of making up to several mg of RNAs longer than about 20nts with relatively high quality as compared to solid phase synthesis.

Jakob Suckale at 14:49, 20 June 2009

Jakob Suckale — Sat, 20 Jun 2009 14:49:42 GMT

←Older revision		Revision as of 14:49, 20 June 2009
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	'''In vitro T7 transcription''' is the synthesis of RNAs using a T7 promoter and purified enzyme. It is the standard method of making up to several mg of RNAs longer than about 20nts with relatively high quality as compared to solid phase synthesis.		'''In vitro T7 transcription''' is the synthesis of RNAs using a T7 promoter and purified enzyme. It is the standard method of making up to several mg of RNAs longer than about 20nts with relatively high quality as compared to solid phase synthesis.

-	T7 transcribed RNAs are important components for many molecular biology experiments. They can be used to generate (antisense) RNA probes for blot hybridisation and nuclease protection assays that are more sensitive than randomly primed DNA probes. Modified nucleotides containing isotopes like <sup>32</sup>P or detectable epitopes like DIG can be integrated into the RNA via T7 transcription. Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments.	+	In vitro transcribed RNAs like those from T7 or similar viral promoters like T3 and SP6 are important components for many molecular biology experiments. They can be used to generate (antisense) RNA probes for blot hybridisation and nuclease protection assays that are more sensitive than randomly primed DNA probes. Modified nucleotides containing isotopes like <sup>32</sup>P or detectable epitopes like DIG can be integrated into the RNA via T7 transcription. Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments.

	== Lab-specific protocols ==		== Lab-specific protocols ==

Jakob Suckale: external links

Jakob Suckale — Sat, 20 Jun 2009 14:45:01 GMT

external links

←Older revision		Revision as of 14:45, 20 June 2009
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	* vectors with an N-terminal T7 sequence: [http://openwetware.org/images/2/27/PET11a.pdf pET11a-d], [http://openwetware.org/images/0/08/Vector_Map_-_pET-3a.pdf pET-3a-d]		* vectors with an N-terminal T7 sequence: [http://openwetware.org/images/2/27/PET11a.pdf pET11a-d], [http://openwetware.org/images/0/08/Vector_Map_-_pET-3a.pdf pET-3a-d]
	* vector for blue/white screening and T7 transcription [http://openwetware.org/images/1/1c/PETBlue_2.pdf pETBlue]		* vector for blue/white screening and T7 transcription [http://openwetware.org/images/1/1c/PETBlue_2.pdf pETBlue]
		+
		+	== External links ==
		+	* [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)]
		+	* [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion]
		+	* [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia]
		+

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:RNA]]		[[Category:RNA]]
	[[Category:In vitro]]		[[Category:In vitro]]

Jakob Suckale: lead section

Jakob Suckale — Sat, 20 Jun 2009 14:38:39 GMT

lead section

←Older revision		Revision as of 14:38, 20 June 2009
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		+	'''In vitro T7 transcription''' is the synthesis of RNAs using a T7 promoter and purified enzyme. It is the standard method of making up to several mg of RNAs longer than about 20nts with relatively high quality as compared to solid phase synthesis.
		+
		+	T7 transcribed RNAs are important components for many molecular biology experiments. They can be used to generate (antisense) RNA probes for blot hybridisation and nuclease protection assays that are more sensitive than randomly primed DNA probes. Modified nucleotides containing isotopes like <sup>32</sup>P or detectable epitopes like DIG can be integrated into the RNA via T7 transcription. Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments.
		+
	== Lab-specific protocols ==		== Lab-specific protocols ==
	* [[Sauer:In vitro transcription with T7 RNA polymerase]]		* [[Sauer:In vitro transcription with T7 RNA polymerase]]

Jakob Suckale: see also

Jakob Suckale — Sat, 20 Jun 2009 14:13:47 GMT

Jakob Suckale: lab-specific protocols

Jakob Suckale — Sat, 20 Jun 2009 14:06:24 GMT

lab-specific protocols

←Older revision		Revision as of 14:06, 20 June 2009
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-	[[Sauer:In vitro transcription with T7 RNA polymerase]]	+	== Lab-specific protocols ==
		+	* [[Sauer:In vitro transcription with T7 RNA polymerase]]
		+	* [[Knight:In vitro transcription]]

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:RNA]]		[[Category:RNA]]
	[[Category:In vitro]]		[[Category:In vitro]]

Jakob Suckale: * page restarted * (after content had been moved to lab-specific page)

Jakob Suckale — Sat, 20 Jun 2009 14:05:46 GMT

*** page restarted *** (after content had been moved to lab-specific page)

←Older revision		Revision as of 14:05, 20 June 2009
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		+
	[[Sauer:In vitro transcription with T7 RNA polymerase]]		[[Sauer:In vitro transcription with T7 RNA polymerase]]

Jenny T Nguyen at 18:59, 10 July 2006

Jenny T Nguyen — Mon, 10 Jul 2006 18:59:14 GMT

←Older revision		Revision as of 18:59, 10 July 2006
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	[[Sauer:In vitro transcription with T7 RNA polymerase]]		[[Sauer:In vitro transcription with T7 RNA polymerase]]
		+
		+	[[Category:Protocol]]
		+	[[Category:RNA]]
		+	[[Category:In vitro]]

←Older revision		Revision as of 14:13, 20 June 2009
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-
	== Lab-specific protocols ==		== Lab-specific protocols ==
	* [[Sauer:In vitro transcription with T7 RNA polymerase]]		* [[Sauer:In vitro transcription with T7 RNA polymerase]]
	* [[Knight:In vitro transcription]]		* [[Knight:In vitro transcription]]
		+
		+	== Related OWW pages ==
		+	* [[Rebuilding T7]]
		+	* [[Endy:Dedicated systems/Transcription]]
		+	* vectors with an N-terminal T7 sequence: [http://openwetware.org/images/2/27/PET11a.pdf pET11a-d], [http://openwetware.org/images/0/08/Vector_Map_-_pET-3a.pdf pET-3a-d]
		+	* vector for blue/white screening and T7 transcription [http://openwetware.org/images/1/1c/PETBlue_2.pdf pETBlue]

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:RNA]]		[[Category:RNA]]
	[[Category:In vitro]]		[[Category:In vitro]]