Difference between revisions of "Making RNA probes with T7 transcription"

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== mRNA sense & antisense probes ==
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[[Image:Antisense_probes_and_mRNA.png|thumb|left|500px|Probes need to be antisense respective to the mRNA to be able to bind their target. Care has to be taken during experimental design to make a suitable antisense probe.]]
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== In vitro T7 transcription of an RNA probe ==
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[[Image:T7 primer design for RNA antisense probes.png|thumb|left|500px|Designing primers for T7 RNA synthesis]]
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# designing a suitable primer combining the T7 promoter sequence and a sequence specific part
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# attaching a T7 promoter via PCR
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# in vitro transcription using T7 polymerase and the PCR product as a template
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== See also ==
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* [[In vitro transcription with T7 RNA polymerase]]
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* [[Primer design]]
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== External links ==
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* [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)]
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* [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion]
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* [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia], [http://www.fermentas.com/catalog/modifyingenzymes/rnapolymerases.htm RNA polymerases - Fermentas]]
  
 
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[[Category:RNA]]
 
[[Category:In vitro]]
 
[[Category:In vitro]]

Latest revision as of 07:33, 20 June 2009

back to protocols

mRNA sense & antisense probes

Probes need to be antisense respective to the mRNA to be able to bind their target. Care has to be taken during experimental design to make a suitable antisense probe.


In vitro T7 transcription of an RNA probe

Designing primers for T7 RNA synthesis


  1. designing a suitable primer combining the T7 promoter sequence and a sequence specific part
  2. attaching a T7 promoter via PCR
  3. in vitro transcription using T7 polymerase and the PCR product as a template


See also

External links