RNA blot (Northern)

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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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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.
For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].
For a schematic overview of the method see [http://www.biochem.arizona.edu/classes/bioc471/pages/Lecture2/AMG1.14-North.gif here].

Revision as of 09:40, 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]

See also

External links

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