Making RNA probes with T7 transcription

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(In vitro T7 transcription of an RNA probe)
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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
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== See also ==
== See also ==

Current revision

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Contents

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.
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 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

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