Description

• NeoSupreme qPCR Green Master Mix (2x), ROX: high-efficiency mix for sensitive, fast, and reproducible qPCR.
• Hot-start enzyme mechanism enhances specificity and sensitivity.
• Contains green intercalating dye, dNTPs, stabilizers, enhancers, and ROX reference dye.
• Designed for accurate gene expression analysis and broad instrument compatibility.
• Ready to use: simply add primers and DNA template.

Storage conditions and reagents preparation

This master mix should be stored at -85°C to -15°C in a freezer without defrost cycles in order to guarantee maximal shelf life. Minimize the number of freeze-thaw cycles by storing in working aliquots. The product will remain stable till the expiry date if stored as specified. The green dye is light sensitive, as such the master mix should be protected from light whenever possible.

Compatible real-time PCR instruments

The master mix is compatible with instruments that measure the passive reference signal. However, it is also compatible with instruments that do not require a passive reference signal for data normalization. The NeoSupreme qPCR Green Master Mix (2x), ROX was optimized to be compatible with the following real-time PCR instruments: Applied Biosystems: 7500; 7500 FAST; QuantStudio™ 5, 6, 7, 12k Flex & ViiA7™

Quality control assay

• Genomic DNA contamination

The product must comply with internal standards of DNA contamination as evaluated through realtime qPCR.

• Nuclease assays

To test for DNase contamination, 0.2-0.3 μg of pNZY28 plasmid DNA are incubated with the master mix for 14-16 h at 37 °C. To test for RNase contamination, 1 μg of RNA is incubated with the master mix for 1 h at 37 °C. Following incubation, the nucleic acids are visualized on a GreenSafe-stained agarose gel. There must be no visible nicking or cutting of the nucleic acids.

• Functional assay

NeoSupreme qPCR Green Master Mixes are extensively tested for activity, processivity, efficiency, sensitivity and heat activation.