NZY First-Strand cDNA Synthesis Kit

NZY First-Strand cDNA Synthesis Kit is a system that includes all the necessary components to synthesize first-strand cDNA, except the template RNA.The resulting single-stranded cDNA is suitable for use in real-time quantitative Reverse Transcription PCR (RT-qPCR). NZY First-Strand cDNA Synthesis Kit 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.

The kit includes a combination of random hexamers and oligo(dT)₁₈ primers in order to increase sensitivity. The primers are included in the NZYRT 2× Master Mix, which also contains dNTPs, MgCl₂ and an optimized RT buffer. NZYRT Enzyme Mix includes both the NZY Reverse Transcriptase (RNase H minus) and the 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 reaction since PCR primers will bind more easily to the cDNA.

Features:

• Provides high yields of full-length cDNA products
• Formulated to increase sensitivity in RT-qPCR
• Primer type: oligo(dT)₁₈ and random hexamers
• 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

High quality cDNA with low levels of starting RNA:
NZY First-Strand cDNA Synthesis Kit is an ideal choice for reverse transcription of low abundant RNA templates. NZYRT Enzyme Mix present in the kit shows high sensitivity, allowing synthesis of a wide range of template amounts.

A 10-fold serial dilution of total RNA from mouse liver (1 μg to 10 pg) was reverse transcribed in a 20 μL reaction using the NZY First-Strand Synthesis Kit. 1 μL of the resultant cDNA was then used as template in an end-point PCR using specific primers for amplification of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene with Supreme NZYTaq DNA polymerase.  NC: No template control. Lane M: NZYDNA Ladder III.

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
• NZY RNase H (E. coli)
• DEPC-treated H₂O

Reverse Transcription kits available at NZYTech: Selection Guide

1. NZY First-Strand cDNA Synthesis Kit, separate oligos
2. NZY M-MuLV First-Strand cDNA Synthesis Kit, separate oligos
3. NZY First-Strand cDNA Synthesis Kit, no oligos
4. NZY M-MuLV First-Strand cDNA Synthesis Kit, no oligos