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Nucleic Acids Research, 1977, Vol. 4, No. 2 339-352
© 1977

Articles

Sequence analysis of T1 ribonuclease fragments of 18S ribosomal RNA by 5'-terminal labeling, partial digestion, and homochromatography fingerprinting

Motohiro Fuke and Harris Busch

Department of Pharmacology, Baylor College of Medicine Houston, TX 77030, USA

Received November 3, 1976. The method employed to determine the sequence of a Tl RNase fragment, A-A-A-A-A-U-A-A-C-A-A-U-A-C-A-Gp, from Novikoff rat hepatoma 18S ribosomal RNA is described. This method is applicable to any oligoribonucleotide produced by specific endonucleases that leave the newly cleaved 5'-end free for labeling with polynucleotide kinase and y-[32p]-ATP. The [32p]-labeled oligoribonucleotide is subjected to partial endonucleolytic digestion and fractionated by two-dimensional homochromatography fingerprinting. The nucleotide sequence is determined by following mobility shifts of the labeled and partially digested oligoribonucleotides in homochromatography fingerprinting.


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