Accuris qMax cDNA Synthesis Kit, 250 reactions

Accuris original cDNA Synthesis Kit comes as a convenient 2-tube format for easy reaction set up and is ideal for 4pg to 0.5µg of input RNA. One tube includes our exceptionally stable Reverse Transcriptase combined with a potent RNAse inhibitor, and the other tube contains a 5X reaction buffer with an optimal mixture of anchored oligo (dT) primers and random hexamers to produce a non-biased population of cDNA.

Accuris™ qMAX cDNA Synthesis Kit
High quality, qPCR ready DNA is quickly generated with the Accuris qMax cDNA Synthesis kit. A blend of hexamer and oligo (dT) primers ensures unbiased and reproducible synthesis with true, relative cDNA representation. The exceptionally thermostable qMax Reverse Transcriptase combined with a potent RNase inhibitor allows for transcription to take place at higher temperatures (alleviating secondary structure problems) and maintains the integrity of the total RNA. The 5X buffer provides the perfect environment for highly efficient cDNA synthesis and greater yields. The buffer is optimized for superior data accuracy and reproducibility with limited starting material and low-copy templates.

• Unbiased, complete sequence representation for pg to 2μg of total RNA
• Thermostable reverse transcriptase and optimized next generation buffer system provide consistent, high yield product

Upon receipt, immediately store at -20°C. Avoid excessive freeze/thaw cycles. When stored as directed, this product will retain its activity for 12 months from date of receipt. The product may also be stored at 4°C for up to one month. Limitations of Use For research purposes only. Not intended for therapeutic or diagnostic use.

Quality Control
Accuris enzymes and reagents are tested under general assay conditions for activity, reproducibility, efficiency, heat activation, sensitivity, and absence of nuclease contamination and nuclease activity. This product is manufactured under a comprehensive quality management system, following ISO 9001:2008 standards. Accuris is not responsible for consequential or incidental damages, direct or indirect, resulting from use of this product. Accuris guarantees the performance of this product as described when used in accordance with these instructions.

General Guidelines
Buffer
The cDNA Synthesis Kit is supplied with a specially formulated 5X buffer. This buffer has been optimized to work with the RT provided in the kit and contains a ratio of anchored oligo (dT) primers and random hexamers determined to produce a nonbiased population of cDNA. High quality cDNA can be produced with maximum efficiency using this buffer and RT. The use of other additives is not required or recommended.
Template
The cDNA Synthesis Kit works optimally with 0.1μg to 4.0pg of total RNA or 2.0μg to 0.2pg of oligo(dT) purified mRNA. The amount of starting material for low copy number genes can be increased up to 2.0μg for better results.
Reaction conditions
Most reactions can be carried out at a temperature of 42 & deg;C for 30 minutes. When working with templates that have a high GC content (above 65%), the temperature can be increased, up to 55 & deg;C, to alleviate any problems associated with secondary structure.
Analysis by qPCR
A cDNA synthesis reaction produces enough cDNA for analysis by real time PCR as well as other down stream processing reactions. For a 20μl qPCR reaction, using 4.0μl of the cDNA synthesis reaction product is recommended.

Package contents and reordering
The Accuris qMax cDNA Synthesis Kit is available in 250-reaction packs. Kit includes 20X reverse transcriptase with RNase inhibitor and 5X reaction buffer with primers.

Protocol
1. Incubate tube from above at 42°C for 30 minutes. For RNA with a high degree of secondary structure, incubate at 55°C.
2. Denature the RT by incubating at 85°C for 10 minutes.
The resulting cDNA can be stored and/or an aliquot can be used for qPCR using Accuris qMAX qPCR products.
Accuris offers a full line of PCR enzymes and master mixes. Use our convenient PCR Selector Tool to determine the correct Accuris PCR Reagent for your applications.

Reaction Setup
Thaw the 5X buffer and prepare your reaction in a 0.2ml tube as below:

For other volumes, adjust the amount of each component accordingly.

Please note: This product is not returnable.