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TranscriptAid™ T7 High Yield Transcription Kit
High yield in vitro transcription

#K0441	for 50 reactions
Related Documents (in pdf, ~50-90 KB): Complete Protocol: #K0441 MSDS (English) MSDS (English-USA) MSDS (German) QuickProtocol™ (in pdf, 73 KB) Brochure (in pdf, 189KB)

 

Features

• Exceptionally high yields of RNA transcripts both long and short.
• Guaranteed integrity of synthesized RNA.
• Milligram amounts of RNA can be generated in a single, scaled-up reaction.
• Flexible format allows production of high quality unlabeled, labeled or capped RNA.
• Superior composition - all necessary reagents included for an entire procedure.

TranscriptAid™ T7 High Yield Transcription Kit, brochure in pdf, 189 KB

Description
The TranscriptAid™ T7 High Yield Transcription Kit is designed for high yield in vitro transcription from DNA templates containing T7 RNA Polymerase promoter. The kit contains reagents for 50 reactions of 20 µl. Depending on transcript length, each reaction yields approximately 150 µg RNA from 1 µg template in 2 hours, 10 times greater than it is achievable in conventional in vitro transcription reactions (Fig.1). The reaction can be scaled-up to produce milligram amounts of full-length RNA.
The kit provides all components for transcription reaction, transcript loading and analysis on a gel. The TranscriptAid™ Enzyme Mix contains T7 RNA polymerase conveniently premixed with recombinant RiboLock™ RNase Inhibitor to ensure integrity of RNA transcript. DNase I, RNase-free, is provided for efficient removal of template DNA after transcription reaction.
The 2X RNA Loading Dye is included for convenience in RNA loading. The RiboRuler™ High Range RNA Ladder, ready-to-use aids in RNA sizing and quantification on a gel. NTPs are provided in individual tubes for flexibility in synthesis of non-radioactively labeled probes
or capped RNA.
The RNA synthesized is suitable in all applications that require large amounts of RNA, such as in vitro translation, antisense RNA and RNAi studies, RNase protection assays, studies of RNA splicing, isolation of RNA binding proteins. Non-radioactively labeled RNA can be used as probes in microarrays, blots or in situ hybridization.
Due to large quantities of RNA synthesized with TranscriptAid™ T7 High Yield Transcription Kit, generation of high specific activity radiolabeled probes would require prohibitively large amounts of radiolabeled nucleotide. Fermentas T7 Transcription Kit is a recommended alternative for this application.

Figure 1. Time course of RNA synthesis. 500 nt, 1000 nt and 6000 nt RNA transcripts were generated with TranscriptAid™ T7 High Yield Transcription Kit. Yields of RNA were determined at different transcription reaction time points using Agilent Bioanalyzer 2100.

Figure 2. Superior integrity of RNA transcripts. 500 nt, 1000 nt and 6000 nt RNA transcripts were generated in 20µl high yield in vitro transcription reactions containing 1µg DNA template in 2h at 37°C. 6µl of 80X diluted transcription reaction products were loaded and run on 1% agarose/TAE gel supplemented with 0.5µg ethidium bromide. M - RiboRuler™ RNA Ladder, High Range, ready-to-use 1, 4, 7 - 500 nt RNA transcripts 2, 5, 8 - 1000 nt RNA transcripts 3, 6, 9 - 6000 nt RNA transcripts

Applications

• Generation of RNA transcripts suitable for:
  • in vitro translation,
  • RNase protection assays,
  • RNA binding protein assays,
  • antisense RNA and RNAi,
  • microinjection studies.
• Generation of hybridization probes for:
  • microarrays,
  • in situ hybridization,
  • blotting,

Quality Control
All components of the kit are tested in a functional control assay. A 20µl reaction mixture containing 1µg of the control template DNA yields 140-170µg of intact RNA after a 2h incubation.

Components of the Kit

• TranscriptAid™ Enzyme Mix
• 5X TranscriptAid™ Reaction Buffer
• DNase I, RNase-free
• ATP Solution
• CTP Solution
• GTP Solution
• UTP Solution
• Control Template
• 3 M Sodium Acetate Solution, pH 5.2
• DEPC-treated Water
• 2X RNA Loading Dye
• RiboRuler™ High Range RNA Ladder, ready-to-use
• 0.5 M EDTA, pH 8.0
• Detailed Protocol (in pdf, ~66 KB):
  • #K0441

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