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The Thermo Scientific TranscriptAid T7 High Yield Transcription Kit contains reagents for 50 reactions of 20 µL. Each reaction yields up to 200 µg RNA from 1 µg of template in 2 hours (see Figures 1 and 2). The reaction can be scaled up to produce milligram amounts of full-length RNA. The kit can be used to produce both long and short transcripts for applications that require large yields of RNA (see Figure 3). All necessary reagents for transcription are included, as well as the RiboRuler High Range RNA Ladder for sizing and quantification.
Figure 1 | Time course of RNA synthesis. 500 b, 1000 b and 6000 b RNA transcripts were generated with TranscriptAid T7 High Yield Transcription Kit. 1 µg of DNA template was used in 20 µL reactions. Yields of RNA were determined at different transcription reaction time points using Agilent 2100 Bioanalyzer®.
Figure 2 | Time course of short RNA synthesis. 50 b and 100 b RNA transcripts were generated with TranscriptAid T7 High Yield Transcription Kit in 0 to 8 hour reaction time. Yields of RNA were determined at different transcription reaction time points using Agilent 2100 Bioanalyzer®.
Figure 3 | High yield of full length RNA transcripts. 500, 1000 and 6000 base RNA transcripts were generated using high yield in vitro transcription reactions containing 1 µg of DNA template. Each 20 µL reaction was incubated for 2 hours at 37°C. The transcription products were diluted 80-fold and 6 µL was run on a 1% agarose/TAE gel supplemented with 0.5 µg/mL ethidium bromide.
M – RiboRuler High Range RNA Ladder, ready-to-use1, 4, 7 – 500 b RNA transcripts2, 5, 8 – 1000 b RNA transcripts3, 6, 9 – 6000 b RNA transcripts