Initiator oligonucleotides for the combination of chemical and enzymatic RNA synthesis
1992; Elsevier BV; Volume: 112; Issue: 1 Linguagem: Inglês
10.1016/0378-1119(92)90309-d
ISSN1879-0038
AutoresChristian Pitulle, Reinhard G. Kleineidam, Brian S. Sproat, Guido Krupp,
Tópico(s)RNA Research and Splicing
ResumoTranscription 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 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.
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