NZY First-Strand cDNA Synthesis Kit, separate oligos, is a system that includes all the necessary components to synthesize first-strand cDNA, except the template RNA. Random hexamers and Oligo(dT)18 primers are provided in separate tubes to offer the convenience to choose the appropriate primer to initiate your reverse-transcription reaction. The resulting single-stranded cDNA is suitable for use in real-time quantitative Reverse Transcription PCR (RT-qPCR). NZY First-Strand cDNA Synthesis Kit, separate oligos, is formulated to provide high yields of full-length cDNA products and to increase sensitivity in RT-qPCR. Starting material can range from 10 pg up to 5 µg of total RNA. Besides random hexamers and Oligo(dT)18 primers, the kit includes NZYRT 2× Master Mix, no oligos, which contains dNTPs, MgCl2 and an optimized RT buffer; 10× Annealing Buffer and NZYRT Enzyme Mix. NZYRT Enzyme Mix includes both NZY Reverse Transcriptase (RNase H minus) and NZY Ribonuclease Inhibitor in order to protect RNA against degradation due to ribonuclease contamination. RNase H (from E. coli) is provided in a separate tube to specifically degrade the RNA template in cDNA-RNA hybrids after the first-strand cDNA synthesis. This procedure will improve the sensitivity of subsequent RT-qPCR reactions since PCR primers will bind more easily to the cDNA.