Making RNA probes with T7 transcription: Difference between revisions
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== mRNA sense & antisense probes == | == 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.]] | [[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 == | == 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]] | [[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 | # designing a suitable primer combining the T7 promoter sequence and a sequence specific part | ||
# attaching a T7 promoter via PCR | # attaching a T7 promoter via PCR | ||
# in vitro transcription using T7 polymerase and the PCR product as a template | # in vitro transcription using T7 polymerase and the PCR product as a template | ||
== See also == | == See also == | ||
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
