Making RNA probes with T7 transcription

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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

• In vitro transcription with T7 RNA polymerase
• Primer design

External links

• T7 transcription protocol by Jonathan Davies of Harvard Uni (1998)
• In vitro transcription basics at Ambion
• T7 RNA polymerase - Wikipedia, RNA polymerases - Fermentas]