Optimization of conditions for labeling the 3' OH end of tRNA using T4 RNA ligase

Biochimie
G Keith

Abstract

For several years most primary structure studies of ribonucleic acids have used the [32P] in vitro post-labeling techniques. We adapted our methods from the literature, and simplified them to make them accessible to any laboratory. These procedures are especially useful for preparation and purification of post labeling enzymes: T4 polynucleotide kinase, T4 RNA ligase and of gamma [32P] ATP. We developed a test tube method for 5' [32P] pCp preparation followed by tRNA labeling with T4 RNA ligase. The parameters for optimal labeling were determined. Labeling of 3.10(6) to 5.10(6) Cerenkov CPM per microgram tRNA are currently obtained.

References

Feb 15, 1974·European Journal of Biochemistry·T LinnéL Philipson
Dec 1, 1965·Analytical Biochemistry·H W SiegelmanB C Turner

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Citations

Jan 1, 1987·Journal of Molecular Evolution·A P ArrigoP F Spahr
Nov 1, 1991·Biochimie·C el AdlouniG Keith
Feb 11, 2000·RNA·F Navarro, P Thuriaux
Feb 25, 1992·Nucleic Acids Research·A PrzykorskaG Dirheimer
Feb 25, 1993·Nucleic Acids Research·C el AdlouniA Przykorska
Aug 15, 1997·Nucleic Acids Research·Z E ZehnerW M Holmes
Aug 15, 1998·Nucleic Acids Research·G PrzewlockiA Przykorska
Apr 15, 2005·PLoS Biology·Stepánka VanácováWalter Keller
Mar 30, 1984·Biochemical and Biophysical Research Communications·G PixaG Keith
Dec 16, 2003·The Journal of Biological Chemistry·Nolan R FilipenkoDavid M Waisman

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