Fig. 3.

Analysis of the tRNA ligation products. (A) tRNA halves from [α-³²P]ATP labeled pre-tRNA was used as substrates for the in vitro RNA ligation assay resulting in ligated tRNA and circular intron. (B) The circular intron formed by the action of the M. kandleri extract, P aerophilum RtcB, or T4 polynucleotide kinase + T4 RNA ligase was recovered from a 12% polyacrylamide/8 M urea gel by passive elution and used for nuclease P1 hydrolysis. The resulting nucleoside 5′-monophosphate mixture was separated by thin-layer chromatography on cellulose plates in solvent A. Position of markers are indicated.