Synthesis of adenosine derivatives as transcription initiators and preparation of 5′ fluorescein- and biotin-labeled RNA through one-step in vitro transcription

Fluorescein-linked adenosine nucleotide-initiated transcription. RNA products were resolved by 8% denaturing PAGE and quantitated by phosphorimaging. Transcription in the absence (lane 1) or presence (2 mM) of fluorescein derivatives (5-isomers) of adenosine nucleotides (lanes 2–13). Transcription initiation efficiency varies from 0% to 65% (indicated under the gel, depending on the linker size and hydrophobicity) under the transcription conditions.