The First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from mRNA or total RNA templates. The kit uses M-MuLV Reverse Transcriptase, which has lower RNase H activity compared to AMW reverse transcriptase. The enzyme maintains activity at 37 °C and is suitable for synthesis of cDNA up to 9 kb. The recombinant Thermo Scientific RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA from degradation at temperatures up to 55 °C.
The kit is supplied with both oligo(dT)18 and random hexamer primers. Random hexamer primers bind non-specifically and are used to synthesize cDNA from all RNAs in total RNA population. The oligo(dT)18 primer selectively anneals to the 3'-end of poly(A) RNA, synthesising cDNA only from poly(A) tailed mRNA. Gene-specific primer may also be used with the kit to prime synthesis from a specified sequence. First strand cDNA synthesised with this system can be directly used as a template in PCR or real-time PCR. It is also ideal for second strand cDNA synthesis or linear RNA amplification. Radioactively and non-radioactively labelled nucleotides can be incorporated into first strand cDNA for use as a probe in hybridisation experiments, including microarrays.
