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. 2002 Dec 16;100(1):80–85. doi: 10.1073/pnas.262609399

Figure 5.

Figure 5

Transcription on immobilized templates. (A) Experimental design for the purification of a functional RNAP II-containing PIC. Promoter-containing templates, immobilized on magnetic beads, were incubated with nuclear extract, and PICs were captured by using a magnetic particle concentrator. (B) Lane 1 shows transcription from WT SL RNA template attached to the magnetic beads. Lanes 2–4 show transcription from the supernatant (Sup), wash, and purified PICs on the WT SL RNA template. (C) Lanes 2, 4, and 6 show the transcripts produced from purified PICs formed on WT, S-Mut, or Δ-Mut SL templates, respectively. Lanes 1, 3, and 5 show residual transcriptional activity in the supernatants. WT SL RNA template (100 ng) was added along with rNTPs and transcription buffer for supernatant assays.