RNA blot (Northern)

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(lab-specific protocols)
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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]
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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]]
== See also ==
== See also ==
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* [[RNA Extraction]]
* [[RNA Extraction]]
* [[RNA electrophoresis]]
* [[RNA electrophoresis]]
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* [[Endy:Northern Blot, 32P End-Labeled Probes]]
 
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* [[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!)
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[[Category:Protocol]]
[[Category:Protocol]]
[[Category:RNA]]
[[Category:RNA]]
[[Category:In vitro]]
[[Category:In vitro]]

Revision as of 10:17, 16 April 2009

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.

For a schematic overview of the method see here.

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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