Pitulle C, Kleineidam R G, Sproat B, Krupp G
Institut für Allgemeine Mikrobiologie, Christian-Albrechts-Universität, Kiel, F.R.G.
Gene. 1992 Mar 1;112(1):101-5. doi: 10.1016/0378-1119(92)90309-d.
Transcription reactions with T7 RNA polymerase were performed in the presence of short oligonucleotides (oligos) with guanosine at the 3'-end. We obtained transcripts which had included these 'initiator oligos' at their 5'-termini. The oligos could contain mixtures of deoxyribo-, ribo-, 2'-O-methylated and biotinylated nucleotides. Only the 3'-terminal guanosine of these oligos was encoded in the template DNA at the transcription start point, in contrast to the remainder of the sequence. This 5'-terminal sequence is variable and eliminates the limitation that transcripts must start with a 5'-terminal guanosine. With a 5'-biotinylated dinucleotide, we obtained end-labeled RNAs suitable for nonradioactive RNA sequencing.
在3'末端带有鸟苷的短寡核苷酸（oligos）存在的情况下，进行了T7 RNA聚合酶的转录反应。我们获得了在其5'末端包含这些“起始寡核苷酸”的转录本。这些寡核苷酸可以包含脱氧核糖核苷酸、核糖核苷酸、2'-O-甲基化核苷酸和生物素化核苷酸的混合物。与序列的其余部分相反，这些寡核苷酸中只有3'末端的鸟苷在转录起始点的模板DNA中编码。这种5'末端序列是可变的，消除了转录本必须以5'末端鸟苷开头的限制。使用5'-生物素化二核苷酸，我们获得了适用于非放射性RNA测序的末端标记RNA。
