Site specific enzymatic cleavage of RNA.
The hybridization of a DNA oligonucleotide a specific tetramer or longer) will direct a cleavage by RNase H (EC 3.1.4.34) to a specific site in RNA. The resulting fragments can then be labeled at their 5' or 3' ends, purified, and sequenced directly. This procedure is demonstrated with two RNA molecules of known sequence: 5.8S rRNA from yeast (158 nucleotides) and satellite tobacco necrosis virus (STNV) RNA (1240 nucleotides).
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004720:Endonucleases / Q000378:metabolism; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides / Q000032:analysis; D012335:RNA, Ribosomal; D012260:Ribonucleases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000032:analysis; D013379:Substrate Specificity; D013950:Thymus Gland / Q000201:enzymology
D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D012335:RNA, Ribosomal; D004720:Endonucleases; D012260:Ribonucleases