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
- 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