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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

12,274 Views

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10:37 min

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May 10, 2018

DOI:

10.3791/57519-v

DOI:
10.3791/57519-v

•

10:37 min
May 10, 2018

•

14 Views
,
1Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute,University of Lethbridge, 2Department of Neuroscience and the Canadian Centre for Behavioral Neuroscience (CCBN),University of Lethbridge, 3Arnie Charbonneau Cancer Institute,University of Calgary

Summary

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Toeprinting aims to measure the ability of in vitro transcribed RNA to form translation initiation complexes with ribosomes under a variety of conditions. This protocol describes a method for toeprinting mammalian RNA and can be used to study both cap-dependent and IRES-driven translation.

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Toeprinting AnalysisTranslation Initiation ComplexMammalian MRNAsIRESRNA PurificationRabbit Reticulocyte LysateRNase InhibitorATPGMP-PNPDTTSpermidine

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Cite this Article

Ross, J. A., Thakor, N. Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs. J. Vis. Exp. (135), e57519, doi:10.3791/57519 (2018).

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