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Co-transcriptional capping in vitro. Transcribing pulled-down G-less templates in the presence of capping enzyme were assayed for radioactive ribonucleotide incorporation into the 5′ RNA Cap (lane 1) or the RNA body (lane 2). The body was labeled using ATP, CTP, and [α-32P]UTP for 5 min. The Cap was labeled with ATP, CTP, and UTP with [α-32P]GTP added for 30 s followed by unlabeled GTP for 4.5 min. The wild type and D3 forms of TFIIB are indicated.
