Preparation of polyribosome aminoacyl-transfer ribonucleic acid from the muscle of chick embryos.
A procedure for preparing polyribosome aminoacyl-tRNA free from contamination by supernatant aminoacyl-tRNA and free amino acids is described. Important features of the procedure are the use of acidic buffers to help protect the amino acid-tRNA linkage and the inclusion of sodium dodecyl sulphate, to inhibit ribonuclease activity. The specific radioactivity of polyribosome aminoacyl-tRNA is high within 30s and reaches a maximum in 2 1/2 min, well ahead of polyribosome peptides which, as described by Herrmann et al. (1971), attain maximum specific radioactivity in about 10 min.
D000604:Amino Acyl-tRNA Synthetases; D000818:Animals; D001665:Binding Sites; D002642:Chick Embryo; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009132:Muscles / Q000032:analysis; D011132:Polyribosomes / Q000032:analysis*; D012313:RNA / Q000302:isolation & purification; D012343:RNA, Transfer / Q000302:isolation & purification*; D013997:Time Factors
D012313:RNA; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases