Making RNA probes with T7 transcription: Difference between revisions
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== mRNA sense & antisense probes == | |||
[[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 == | |||
[[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 | |||
# attaching a T7 promoter via PCR | |||
# in vitro transcription using T7 polymerase and the PCR product as a template | |||
== See also == | |||
* [[In vitro transcription with T7 RNA polymerase]] | |||
* [[Primer design]] | |||
== External links == | == External links == | ||
* [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)] | * [http://genetics.mgh.harvard.edu/szostakweb/protocols/transcription/index.html T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)] | ||
* [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion] | * [http://www.ambion.com/techlib/basics/transcription/index.html In vitro transcription basics at Ambion] | ||
* [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia] | * [http://en.wikipedia.org/wiki/T7_RNA_polymerase T7 RNA polymerase - Wikipedia], [http://www.fermentas.com/catalog/modifyingenzymes/rnapolymerases.htm RNA polymerases - Fermentas]] | ||
[[Category:Protocol]] | [[Category:Protocol]] | ||
[[Category:RNA]] | [[Category:RNA]] | ||
[[Category:In vitro]] | [[Category:In vitro]] | ||
Latest revision as of 08:33, 20 June 2009
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mRNA sense & antisense probes
In vitro T7 transcription of an RNA probe
- designing a suitable primer combining the T7 promoter sequence and a sequence specific part
- attaching a T7 promoter via PCR
- in vitro transcription using T7 polymerase and the PCR product as a template
