RNA blot (Northern) - Revision history

Jakob Suckale: lead section

Jakob Suckale — Thu, 16 Apr 2009 14:19:44 GMT

lead section

←Older revision		Revision as of 14:19, 16 April 2009
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	The '''RNA blot''' or '''Northern blot''' (named after the [[DNA blot (Southern)]] for genomic DNA fragments) is a molecular biology technique used to separate and identify pieces of RNA. RNA molecules are separated by mass on a gel, transferred (blotted) onto a cellulose or nylon membrane, and then labelled with complementary DNA or RNA molecules. These probes are either radioactive, typically <sup>32</sup>P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.		The '''RNA blot''' or '''Northern blot''' (named after the [[DNA blot (Southern)]] for genomic DNA fragments) is a molecular biology technique used to separate and identify pieces of RNA. RNA molecules are separated by mass on a gel, transferred (blotted) onto a cellulose or nylon membrane, and then labelled with complementary DNA or RNA molecules. These probes are either radioactive, typically <sup>32</sup>P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.

-	~~For a schematic overview~~ of the method ~~see~~ [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif ~~here~~].	+	The principle of the method is nicely illustrated in this [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif diagram] and this [http://www.youtube.com/watch?v=KfHZFyADnNg&NR=1 video].

	== Designing RNA probes ==		== Designing RNA probes ==

Jakob Suckale: lab-specific protocols

Jakob Suckale — Thu, 16 Apr 2009 14:17:00 GMT

lab-specific protocols

←Older revision		Revision as of 14:17, 16 April 2009
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	* minimum probe length around 25 nt (anybody has a reference for this?) [http://www.protocol-online.org/biology-forums/posts/16736.html]		* minimum probe length around 25 nt (anybody has a reference for this?) [http://www.protocol-online.org/biology-forums/posts/16736.html]
	* DNA probes may be usable for both [[qRT-PCR]] and RNA blots [http://www.protocol-online.org/biology-forums/posts/17381.html]		* DNA probes may be usable for both [[qRT-PCR]] and RNA blots [http://www.protocol-online.org/biology-forums/posts/17381.html]
		+
		+	== Lab-specific protocols ==
		+	* [[Endy:Northern Blot, 32P End-Labeled Probes]]
		+	* [[Endy:Northern blot, AlkPhos end-labeled probes]]

	== See also ==		== See also ==
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	* [[RNA Extraction]]		* [[RNA Extraction]]
	* [[RNA electrophoresis]]		* [[RNA electrophoresis]]
-	* [[Endy:Northern Blot, 32P End-Labeled Probes]]
-	* [[Endy:Northern blot, AlkPhos end-labeled probes]]
	* [[BE.109:Systems engineering/Measuring DNA, RNA, protein]]		* [[BE.109:Systems engineering/Measuring DNA, RNA, protein]]

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	* [[Image:1stars.png]] [http://www.roche-applied-science.com/pack-insert/2039672a.pdf Pack insert DIG RNA detection kit], Roche		* [[Image:1stars.png]] [http://www.roche-applied-science.com/pack-insert/2039672a.pdf Pack insert DIG RNA detection kit], Roche
	* [http://www.mcb.arizona.edu/parker/PROTOCOLS/NorthernOligoHyb.htm Probe hybridization protocol] - Parker lab (don't know how many stars to give it!)		* [http://www.mcb.arizona.edu/parker/PROTOCOLS/NorthernOligoHyb.htm Probe hybridization protocol] - Parker lab (don't know how many stars to give it!)
		+
	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:RNA]]		[[Category:RNA]]
	[[Category:In vitro]]		[[Category:In vitro]]

Jakob Suckale: Southern link

Jakob Suckale — Thu, 16 Apr 2009 13:40:06 GMT

Southern link

←Older revision		Revision as of 13:40, 16 April 2009
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-	The '''RNA blot''' or '''Northern blot''' (named after the [[~~Southern~~ blot]] for DNA fragments) is a molecular biology technique used to separate and identify pieces of RNA. RNA molecules are separated by mass on a gel, transferred (blotted) onto a cellulose or nylon membrane, and then labelled with complementary DNA or RNA molecules. These probes are either radioactive, typically <sup>32</sup>P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.	+	The '''RNA blot''' or '''Northern blot''' (named after the [[DNA blot (Southern)]] for genomic DNA fragments) is a molecular biology technique used to separate and identify pieces of RNA. RNA molecules are separated by mass on a gel, transferred (blotted) onto a cellulose or nylon membrane, and then labelled with complementary DNA or RNA molecules. These probes are either radioactive, typically <sup>32</sup>P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.

	For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].		For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].

Barry Canton at 21:08, 4 April 2009

Barry Canton — Sat, 04 Apr 2009 21:08:17 GMT

←Older revision		Revision as of 21:08, 4 April 2009
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-	~~#REDIRECT~~ [[Northern blot]]	+	The '''RNA blot''' or '''Northern blot''' (named after the [[Southern blot]] for DNA fragments) is a molecular biology technique used to separate and identify pieces of RNA. RNA molecules are separated by mass on a gel, transferred (blotted) onto a cellulose or nylon membrane, and then labelled with complementary DNA or RNA molecules. These probes are either radioactive, typically <sup>32</sup>P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.
		+
		+	For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].
		+
		+	== Designing RNA probes ==
		+	* DNA probes, esp. using DIG-antibody detection, often give no/weak signal; RNA probes often better here [http://www.bio.net/bionet/mm/methods/2000-October/085274.html]
		+	* minimum probe length around 25 nt (anybody has a reference for this?) [http://www.protocol-online.org/biology-forums/posts/16736.html]
		+	* DNA probes may be usable for both [[qRT-PCR]] and RNA blots [http://www.protocol-online.org/biology-forums/posts/17381.html]
		+
		+	== See also ==
		+	* [[RNA]]
		+	* [[RNA Extraction]]
		+	* [[RNA electrophoresis]]
		+	* [[Endy:Northern Blot, 32P End-Labeled Probes]]
		+	* [[Endy:Northern blot, AlkPhos end-labeled probes]]
		+	* [[BE.109:Systems engineering/Measuring DNA, RNA, protein]]
		+
		+	== External links ==
		+	* [[Image:3stars.png]] [http://cancer.ucsd.edu/howelllab/Northern.html RNA blot protocol for cells in culture, complete with materials] (1999) by Howell lab, UCSD
		+	* [[Image:3stars.png]] [http://www.bio.net/bionet/mm/methods/2000-October/085274.html detailed RNA blot protocol for DIG probes] (2000) by Arnoud van Vliet
		+	* [[Image:2stars.png]] [http://www.protocol-online.org/cgi-bin/htsearch?config=htdig_pol&restrict=&exclude=&words=northern+blot archived protocol conversations from Protocol Online]
		+	* [[Image:2stars.png]] [http://www.protocol-online.org/prot/Molecular_Biology/RNA/Northern_Blotting/ index of RNA blot protocols from Protocol Online]
		+	* [[Image:2stars.png]] [http://kellylab.wustl.edu/protocols/RNA_and_northern_blot3-3-2006.pdf RNA preparation and blotting protocol] (2006) by Kelly lab, Washington Uni
		+	* [[Image:2stars.png]] [http://biology.semo.edu/agathman/research/protocols/northern.htm RNA blot protocol] by Allen Gathman from Southeast Missouri State Uni
		+	* [[Image:2stars.png]] [http://www.ambion.com/techlib/basics/northerns/ The basics of RNA blots], Ambion general article
		+	* [[Image:1stars.png]] [http://www.ambion.com/techlib/tn/63/631.html Techniques to detect mRNA - includes RNA blot], Ambion TechNotes
		+	* [[Image:1stars.png]] [http://www.roche-applied-science.com/pack-insert/2039672a.pdf Pack insert DIG RNA detection kit], Roche
		+	* [http://www.mcb.arizona.edu/parker/PROTOCOLS/NorthernOligoHyb.htm Probe hybridization protocol] - Parker lab (don't know how many stars to give it!)
		+	[[Category:Protocol]]
		+	[[Category:RNA]]
		+	[[Category:In vitro]]

Barry Canton: RNA blot (Northern) moved to Northern blot

Barry Canton — Tue, 05 Aug 2008 03:15:07 GMT

RNA blot (Northern) moved to Northern blot

New page

#REDIRECT [[Northern blot]]