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. 2000 Jun 15;28(12):e59. doi: 10.1093/nar/28.12.e59

Figure 4.

Figure 4

Monitoring of in vitro transcription reactions by fluorescence spectral changes. (A–C) A 600-fold excess of the probe pair for the 566–585 site of the xelf1-α RNA was added to the in vitro transcription reaction solution and the reaction carried out. A vector containing xelf1-α cDNA was used as template. (A) Changes in fluorescence spectra during the reaction. Fluorescence spectra at 0, 10, 20, 30, 60, 90, 120 and 180 min are shown. (B) Ratios of fluorescence intensity (I670 nm/I514 nm) of the spectra in (A) plotted against incubation time. (C) Time course of xelf1-α RNA synthesis. At each incubation time an aliquot was removed from the reaction solution and the synthesized RNA extracted. The amounts of RNA were estimated by taking 1 absorbance unit at 260 nm as equivalent to 35 µg. (D–F) In vitro transcription reactions were carried out as described in (A)–(C) using a vector containing human c-fos cDNA instead of the vector containing xelf1-α cDNA as template. (D) Changes in fluorescence spectra during the reaction. Fluorescence spectra at 0, 30, 60 and 180 min are shown. (E) Ratios of fluorescence intensity (I670 nm/I514 nm) of spectra in (D) plotted against incubation time. (F) Time course of c-fos RNA synthesis.