Figure 3.
Synthesis of 17 nt RNA by in vitro transcription with 7-EAATP and T7 RNA polymerase. A) T7 RNA polymerase transcription template for synthesis of 17 nt RNA containing one 7-EAA or A residue (X). B) Products of T7 polymerase transcriptions with template shown in (A) containing 5 mM ATP or 7-EAATP, α -32P GTP and allowed to proceed for 2 h at 37 °C. (arrow indicates major transcription product). C) CuAAC reaction with 17 nt RNAs. Lanes 1-3: product from transcription with 7-EAATP, Lanes 4-6: product from transcription with ATP. Lanes 1 and 4: no CuAAC reaction reagents added, Lanes 2 and 5: CuAAC reagents lacking azide, Lanes 3 and 6: CuAAC reagents plus azide. (*) indicates position of CuAAC product.