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. 1997 May 1;25(9):1862–1863. doi: 10.1093/nar/25.9.1862

Rapid selection of aminoacyl-tRNAs based on biotinylation of alpha-NH2 group of charged amino acids.

J Pütz 1, J Wientges 1, M Sissler 1, R Giegé 1, C Florentz 1, A Schwienhorst 1
PMCID: PMC146665  PMID: 9162902

Abstract

A rapid selection procedure to separate low amounts of aminoacylated tRNAs from large pools of inactive variants is described. The procedure involves a three-step protocol. After initial aminoacylation of a tRNA pool, N-hydroxysuccinimide ester chemistry is applied to biotinylate the alpha-NH2 group of the amino acid bound to the 3'-end of a tRNA. The biotin tag is used to capture the derivatized tRNAs on streptavidin-conjugated magnetic beads. Variants bound to the solid phase can be amplified by RT-PCR and transcription, providing tRNAs for subsequent selection rounds.

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Selected References

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