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Figure 2 | Plant Methods

Figure 2

From: Reverse protection assay: a tool to analyze transcriptional rates from individual promoters

Figure 2

Validation of the Reverse RNase Protection Assay. The 5'-ends of rrn16 (A), rbcL (B), and psbA (C) transcripts were analyzed by RNase Protection Assay (RPA, lanes 2) and Reverse RNase Protection Assay (RePro, lanes 3). Mapped transcription initiation sites (filled circles) are identified by their distance between the transcription initiation site and the first nucleotide of the mature RNA (rrn16) or the translation initiation codon (rbcL, psbA) in nucleotides. Molecular weight marker in nt is provided on the sides (M, lanes 1). The positions of probes, primers, and protected fragments are shown in (D), (E), and (F) for rrn16, rbcL, and psbA, respectively. Scissor symbols denote RNA processing sites. Black boxes denote coding regions; small arrows indicate primers; grey boxes and angled arrows indicate promoters and transcription initiation sites; ball-ended arrows indicate complementary RNA probes; bar-ended lines denote protected RNA fragments. Respective sizes are given in nucleotides (nt). Note that the RNA probe sizes include 6 nt added to the transcripts from the T7 promoter. A scale is provided on the bottom left.

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