The GScriptULTRA First-Strand Synthesis Kit (Oligo-dT) provides a convenient and sensitive of cDNA synthesis from RNA molecules by reverse transcription (RT). The kit consists of five major components: GScriptULTRA RTase, 5X 1st strand buffer, 0.1 M DTT, 50 μM Oligo(dT)₂₀, and Random Hexamers primer, and 10 mM dNTP (Deoxyribonucleotide triphosphates) mix. The GScriptULTRA RTase is a recombinant Moloney Murine Leukemia Virus (M-MLV) transcription polymerase expressed in E. coli and purified to homogeneity. It has lower RNase H activity and high thermal stability. The enzyme is widely used to synthesize first-strand cDNA at temperatures up to 55°C with increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. It can generate cDNA from 100 base pairs (bps) to 12 Kilo base pairs (kb). The 5X Reaction Mix buffer is optimized for reverse transcription. The DTT breaks the disulfide bonds, loosen the secondary structure of RNA, and helps in initiation for cDNA synthesis. The Oligo(dT)₂₀ consist of a stretch of 20 deoxythymidines that anneal to poly(A) tails of
eukaryotic mRNAs. The primers are optimized for constructing cDNA libraries. The dNTP solution consists all four nucleotides (dATP, dCTP, dGTP, dTTP), suitable for use in cDNA synthesis.
- Downstream application for PCR to detection of expressed genes, examination of transcript variants, or generation of cDNA templates for cloning and sequencing.
The quality of the GScriptULTRA First-Strand Synthesis Kit (Oligo-dT) is tested on a lot-to-lot basis to ensure consistent product quality.