Northern blot

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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.
 
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For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].
 
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== Designing RNA probes ==
 
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* 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]
 
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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]
 
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* DNA probes may be usable for both [[qRT-PCR]] and RNA blots [http://www.protocol-online.org/biology-forums/posts/17381.html]
 
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== See also ==
 
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* [[RNA]]
 
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* [[RNA Extraction]]
 
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* [[RNA electrophoresis]]
 
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* [[Endy:Northern Blot, 32P End-Labeled Probes]]
 
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* [[Endy:Northern blot, AlkPhos end-labeled probes]]
 
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* [[BE.109:Systems engineering/Measuring DNA, RNA, protein]]
 
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== External links ==
 
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* [[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
 
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* [[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
 
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* [[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]
 
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* [[Image:2stars.png]] [http://www.protocol-online.org/prot/Molecular_Biology/RNA/Northern_Blotting/ index of RNA blot protocols from Protocol Online]
 
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* [[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
 
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* [[Image:2stars.png]] [http://biology.semo.edu/agathman/research/protocols/northern.htm RNA blot protocol] by Allen Gathman from Southeast Missouri State Uni
 
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* [[Image:2stars.png]] [http://www.ambion.com/techlib/basics/northerns/ The basics of RNA blots], Ambion general article
 
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* [[Image:1stars.png]] [http://www.ambion.com/techlib/tn/63/631.html Techniques to detect mRNA - includes RNA blot], Ambion TechNotes
 
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* [[Image:1stars.png]] [http://www.roche-applied-science.com/pack-insert/2039672a.pdf Pack insert DIG RNA detection kit], Roche
 
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* [http://www.mcb.arizona.edu/parker/PROTOCOLS/NorthernOligoHyb.htm Probe hybridization protocol] - Parker lab (don't know how many stars to give it!)
 
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[[Category:Protocol]]
 
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[[Category:RNA]]
 
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[[Category:In vitro]]
 

Revision as of 16:07, 4 April 2009

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