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Overview of mRNA synthesis work flow with the HiScribe T7 ARCA mRNA Kit ..

HiScribe™ T7 High Yield RNA Synthesis Kit | NEB

Transcriptor Reverse Transcriptase – the core component of the kit – is a recombinant reverse transcriptase expressed in . The enzyme has RNA-directed DNA polymerase activity, DNA-dependent DNA polymerase activity, unwinding activity, and RNase H activity that degrades RNA in RNA:DNA hybrids. The latter circumvents the need to perform an additional time-consuming RNase H incubation step after reverse transcription. This shortens the reaction time and reduces costs.
Transcriptor Reverse Transcriptase is recommended for RT-PCR because of its high sensitivity in combination with high thermostability: the enzyme synthesizes long cDNA products (up to 14 kb) and can be used at temperatures up to +65°C. Due to its thermostability, Transcriptor Reverse Transcriptase is recommended for GC-rich templates with high secondary structure, without the need to include additives in the reaction.
The kit provides all reagents required for first-strand cDNA synthesis reactions. For priming, three different primer systems can be used. Two cDNA synthesis primers are provided with the kit: random hexamer primers and an anchored-oligo(dT)18 primer. The latter is designed to bind at the beginning of the poly(A) tail to generate full-length cDNAs and to prevent priming from internal sites of the poly(A) tail. The 5' ends of long mRNAs are often underrepresented; therefore, this priming method is preferred for most applications. The use of random hexamer primers enables priming throughout the length of RNA for uniform representation of all RNA sequences and allows reverse transcription of RNAs that do not carry a poly(A) tail.
Thermostable Protector RNase Inhibitor is included in the kit to protect RNA from degradation at high reaction temperatures.

mRNAs synthesized withthe kit can be used for cell transfection, microinjection, in vitro translation andRNA vaccines.

HiScribe™ T7 ARCA mRNA Kit | NEB

For efficient translation to occur, most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure at the 5′ end and a poly(A) tail at the 3′ end. The HiScribe™ T7 ARCA mRNA Synthesis Kit () is designed to synthesize capped and tailed mRNAs for variety of applications. Capped mRNAs are synthesized by co-transcriptional incorporation of Anti-Reverse Cap Analog, ARCA, using T7 RNA Polymerase. A poly(A) tail is then added by E. coli Poly(A) Polymerase. This kit is also available without E. coli Poly(A) Polymerase (NEB #E2065S) for use with DNA templates encoding a poly(A) stretch or not requiring a poly(A) tail. Both kits include DNase I and LiCl for DNA template removal and quick mRNA purification.

HiScribe™ T7 Quick High Yield RNA Synthesis Kit; HiScribe™ T7 ARCA mRNA Kit …

For efficient translation to occur, most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure at the 5′ end and a poly(A) tail at the 3′ end. The HiScribe™ T7 ARCA mRNA Synthesis Kit () is designed to synthesize capped and tailed mRNAs for variety of applications. Capped mRNAs are synthesized by co-transcriptional incorporation of Anti-Reverse Cap Analog, ARCA, using T7 RNA Polymerase. A poly(A) tail is then added by E. coli Poly(A) Polymerase. This kit is also available without E. coli Poly(A) Polymerase (NEB #E2065S) for use with DNA templates encoding a poly(A) stretch or not requiring a poly(A) tail. Both kits include DNase I and LiCl for DNA template removal and quick mRNA purification.

biotechrabbit™ cDNA Synthesis Kit provides superior components that ensure efficient first-strand cDNA synthesis from mRNA or total RNA templates