RNAMaxx High Yield Transcription Kit - Details & Specifications

Traditional transcription methods often produce RNA yields that are insufficient for many downstream applications. The RNAMaxx High Yield Transcription Kit has been designed to produce large quantities of RNA for many gene expression analysis applications including RNA interference, RNase protection assays, antisense or microinjection studies, and the study of RNA binding proteins. Modified ribonucleotides are easily incorporated into the transcripts for use in constructing non-isotopically labeled RNA probes for hybridization studies or microarrays. The RNAMaxx transcription kit produces 80 to 100 µg or more of RNA from 1 µg of DNA template in just 2 hours. In addition to producing large quantities of RNA, the RNAMaxx transcription kit easily produces transcripts from a wide range of DNA template sizes, up to 9 kb.

The RNAMaxx transcription kit uses T7 RNA polymerase to synthesize RNA from a double-stranded DNA template containing the T7 promoter. To generate high RNA yields, high concentrations of ribonucleotides are added to the reaction mixture. When a ribonucleotide is incorporated into RNA, a molecule of pyrophosphate is released. High concentrations of pyrophosphate have been shown to inhibit further polymerization. To remove this inhibition, yeast inorganic pyrophosphatase has been added to the reaction. The use of a higher concentration of ribonucleotides and the addition of yeast inorganic pyrophosphatase in the reaction mixture increases the RNA yield 10 to 30 times the yield produced by traditional in vitro transcription methods.

The RNAMaxx high yield transcription kit is designed to produce large quantities of RNA quickly and easily from a DNA template. Just 1 µg of template can yield 80 to 100 µg or more of RNA transcript in one short incubation. This is 10 to 30 times the yield produced by a traditional in vitro transcription reaction. The kit uses T7 RNA polymerase to produce high yields of RNA in just two hours. Figure 1 compares the RNAMaxx high yield transcription kit with in vitro transcription kits from two competitors. Using a 1.8-kb DNA template we found that after two hours, higher yields of RNA were obtained using the RNAMaxx transcription kit.

The RNAMaxx high yield transcription kit is ideal for the construction of non-isotopically labeled RNA probes. A variety of modified nucleotides including amino allyl- and biotin-labeled nucleotides are easily incorporated using the kit. The resulting labeled RNA transcripts can be used as probes for microarrays, Northern blot analysis, or in situ hybridization.

The RNAMaxx high yield transcription kit is ideal for the construction of non-isotopically labeled RNA probes. A variety of modified nucleotides including amino allyl- and biotin-labeled nucleotides are easily incorporated using the kit. The resulting labeled RNA transcripts can be used as probes for microarrays, Northern blot analysis, or in situ hybridization.

In addition to producing large quantities of RNA, the RNAMaxx high yield transcription kit easily produces transcripts from a wide variety of template sizes, as illustrated in Figure 2 below. Samples of RNA transcribed from three template lengths, 1.8 kb, 4 kb and 9 kb, were analyzed in parallel by gel electrophoresis and by Agilent 2100 Bioanalyzer. RNA yields were then quantified using fluorescent staining in Figure 3. Virtually identical yields of near 100 µg were observed with each template size, demonstrating both the consistency and the versatility of the RNAMaxx high yield transcription kit.

For Research Use Only. Not for use in diagnostic procedures