NZY First-Strand cDNA Synthesis Kit, separate oligo

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.

The 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

Features:

• Possibility to choose the primer to initiate the reaction
• Provides high yields of full-length cDNA products
• Formulated to increase sensitivity in RT-qPCR
• Starting material: 10 pg to 5 µg of total RNA
• Optimal reaction temperature: 50 °C
• Convenient and reliable

Applications:

• First-strand cDNA synthesis for use in real-time quantitative RT-PCR (RT-qPCR)
• Two-step RT-PCR assays
• Ideal for high GC-content templates due to the optimum activity at 50 ºC

Specifications:

• Storage conditions: Store all kit components at -20ºC
• Shipping conditions: Shipped at 4 ºC to dry ice

Components:

• NZYRT Enzyme Mix
• NZYRT 2x Master Mix, no oligos
• 10× Annealing Buffer
• Random hexamer mix (50 ng/µL)
• Oligo(dT)18 primer mix (50 µM)
• NZY RNase H (E. coli)
• DEPC-treated H2O