说明

The RevertAid™ Premium First Strand cDNA Synthesis Kit is a complete system for highly efficient synthesis of first strand cDNA. The kit uses the RevertAid™ Premium Reverse Transcriptase (RT), an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features the highest thermostability among the derivatives of M-MuLV RT and lacks the RNase H activity. The RevertAid™ Premium First Strand cDNA Synthesis Kit allows synthesis of long cDNA up to 20 kb at elevated temperature (up to 60°C) superceeding other systems in ability to prepare full length cDNA. Due to increased synthesis rates of the enzyme first strand cDNA synthesis, the reaction can be completed in 30 min.

RevertAid™ Premium Enzyme Mix contains RevertAid™ Premium Reverse Transcriptase and RiboLock™ RNase Inhibitor. RiboLock™ RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.

Oligo(dT)₁₈ and random hexamer primers are supplied with the kit. Random hexamer primers bind non-specifically and are used to synthesize cDNA from all RNAs in a total RNA population. The oligo(dT)₁₈ primer selectively anneals to the 3’-end of poly(A) RNA, synthesizing cDNA only from poly(A) tailed mRNA. Gene-specific primers may also be used with the kit to prime synthesis from a specified sequence.

Water, nuclease-free is provided for reaction set-up and dilution of sample DNA. The absence of endo-, exodeoxyribonucleases, ribonucleases and phosphatases has been confirmed by appropriate quality tests.

质量控制

Functionally tested in RT-PCR using 100 fg of control GAPDH RNA and GAPDH control primers generate a prominent 496 bp product on 1% agarose gel after ethidium bromide staining.

组分

• RevertAid™ Premium Enzyme Mix
• 5X RT Buffer
• dNTP Mix, 10 mM each
• Oligo(dT)₁₈ Primer
• Random Hexamer Primer
• Water, nuclease-free
• Detailed protocol