Donis-Keller H
Nucleic Acids Res. 1979 Sep 11;7(1):179-92. doi: 10.1093/nar/7.1.179.
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).
DNA 寡核苷酸(特定的四聚体或更长)的杂交将引导核糖核酸酶 H(EC 3.1.4.34)切割 RNA 中的特定位点。然后,所得片段可在其 5' 或 3' 末端进行标记、纯化并直接测序。用两个已知序列的 RNA 分子证明了该程序:来自酵母的 5.8S rRNA(158 个核苷酸)和烟草卫星坏死病毒(STNV)RNA(1240 个核苷酸)。