RNA blot (Northern): Difference between revisions
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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]] | ||
* [[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]] | ||
Revision as of 07:17, 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.
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
RNA blot protocol for cells in culture, complete with materials (1999) by Howell lab, UCSD- detailed RNA blot protocol for DIG probes (2000) by Arnoud van Vliet
archived protocol conversations from Protocol Online- index of RNA blot protocols from Protocol Online
- RNA preparation and blotting protocol (2006) by Kelly lab, Washington Uni
- RNA blot protocol by Allen Gathman from Southeast Missouri State Uni
- The basics of RNA blots, Ambion general article
Techniques to detect mRNA - includes RNA blot, Ambion TechNotes- Pack insert DIG RNA detection kit, Roche
- Probe hybridization protocol - Parker lab (don't know how many stars to give it!)
