The SuperScript III First-Strand Synthesis SuperMix is an optimized formulation for first-strand cDNA synthesis from purified poly(A)⁺ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 0.1 pg to 5 μg of total RNA.

This kit includes SuperScript III/RNaseOUT enzyme mix, 2X First-Strand reaction mix, and an annealing buffer. SuperScript III Reverse Transcriptase is a version of MMLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 45–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than many previous reverse transcriptases.

Using SuperScript III First-Strand Synthesis SuperMix
SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA and can be used to synthesize cDNA from total RNA. RNaseOUT Recombinant RNase Inhibitor is included in the enzyme mix to safeguard against degradation of target RNA due to ribonuclease contamination. The 2X First-Strand reaction mix includes 10 mM MgCl₂ and 1 mM of each dNTP in a buffer formulation that has been optimized for first-strand synthesis of cDNA. The annealing buffer is used in the initial template-primer annealing step. Separate tubes of oligo(dT)₂₀ and random hexamers are also provided. cDNA synthesis can be performed using either total RNA or poly(A)⁺-selected RNA primed with oligo(dT), random primers, or a gene-specific primer.