Life Technologies
5x MEGAscript® T7 Kit
AM1334-5
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In vitro transcription of RNA transcripts has become an important laboratory technique for many cell and molecular biology experiments. The synthesized RNA is used for several downstream applications such as blot hybridization, nuclease protection assays, RNA interference etc. The general aim of the research was to study cell homeostasis during harmful conditions. In particular, the aim of the study was to study different gene functions. And in doing so, the kit was employed to synthesize double stranded RNA for RNA interference, a valuable research tool. The quality and quantity of dsRNA produced using the kit was satisfactory when compared to other In vitro transcription kits.
Gene Knockdown/gene expression studies
Purified PCR product with T7 promoter sites
The manufacturer’s protocol is simple and straightforward to follow. The kit employs DNA template-directed synthesis of RNA. The kit components consists of 10x reaction buffer, enzyme mix, NTPs, nuclease-free water, DNase, control template, ammonium acetate stop solution. The first step is to synthesize or generate the template that contains T7 promoter sites. The reagents of the kit are then used for In vitro transcription using the engineered template. The resulting RNA transcripts are purified via Phenol:chloroform extraction. The purified RNA can be quantified through spectrophotometer and/or by running them on an agarose gel.
It will be helpful to optimize the amount of template and incubation time to obtain optimal RNA yields.
True to its name, Megascript kit does yield large mass amounts of RNA transcripts (See Figure).
None
The kit yields abundant RNA quantities
Pricy
Simple, efficient, worth the money spent on the kit.
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