RNA blot (Northern)

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#REDIRECT [[Northern blot]]
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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.
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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].
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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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== Lab-specific protocols ==
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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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== See also ==
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* [[RNA]]
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* [[RNA Extraction]]
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* [[RNA electrophoresis]]
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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]]

Current revision

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 32P, or contain labelled nucleotides, e.g. DIG-dNTPs, recognisable by antibodies. RNA molecules can be detected and roughly quantified via probe hybridisation.

The principle of the method is nicely illustrated in this diagram and this video.

Contents

Designing RNA probes

  • DNA probes, esp. using DIG-antibody detection, often give no/weak signal; RNA probes often better here [1]
  • minimum probe length around 25 nt (anybody has a reference for this?) [2]
  • DNA probes may be usable for both qRT-PCR and RNA blots [3]

Lab-specific protocols

See also

External links

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