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University of Alabama Huntsville

1 ARTICLES PUBLISHED IN JoVE

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Biology

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
Nitin Shirole 1, Sreeram Balasubramanian 1, Charles Yanofsky 2, Luis Cruz-Vera 1
1Department of Biological Sciences, University of Alabama Huntsville, 2Department of Biology, Stanford University

A major impediment to biochemical analyses of ribosomes containing nascent peptidyl-tRNAs has been the presence of other ribosomes in the same samples, ribosomes not involved in the translation of the specific mRNA sequence being analyzed. We developed a simple methodology to purify, exclusively, the ribosomes containing the nascent peptidyl-tRNA of interest.

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