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!)