The MEGAscript T7 Kit is an ultra-high yield in vitro transcription kit. The high yields are achieved by modifying typical transcription reaction conditions so that very high nucleotide concentrations can be effectively used. The T7 Enzyme Mix and the 10X Reaction Buffer are specifically calibrated for each lot and RNA polymerase.
Using the MEGAscript T7 Kit The MEGAscript T7 Kit contains in vitro transcription reaction components and a control template. The kit will yield a total of 3–5 mg of RNA (approximately 100 µg of RNA or more per reaction) from the control template supplied with the kit. This corresponds to 400–650 moles of RNA for each mole of template. Smaller templates typically yield a lower mass and a higher molar yield of product. Novel transcription reaction conditions and a patented, high-yield technology allow the synthesis of 10–50 times the amount of RNA produced by conventional transcription reactions.
Applications The MEGAscript T7 Kit is intended for the synthesis of large amounts of unlabeled or low specific activity RNA for a variety of uses, including in vitro translation, antisense/microinjection studies, and isolation of RNA binding proteins. In large-scale transcription reactions, the concentration of all 4 nucleotides is high, well above the Km for the enzyme. The MEGAscript T7 Kit typically yields over 10 times more RNA than conventional in vitro transcription reactions. The kit is not recommended for synthesis of high specific activity probes.
Accessory products MessageAmp aRNA amplification kits are powered by MEGAscript technology and have been developed especially for the amplification of cRNA prior to array analysis. To purify in vitro transcription products from free nucleotides, buffer components, and enzymes, Life Technologies recommends using the MEGAclear Kit (Cat. No. AM1908) and the MEGAclear-96 Kit (Cat. No. AM1909).
2