biotechrabbit™ CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus is a ready-to-use, four-times-concentrated formulation that includes an adjusted concentration of heat-labile Uracil-DNA Glycosylase (HL-UDG), minimizing the risk of carryover contamination.

The master mix is optimized for one-step real-time RT-PCR quantification of RNA templates, including mRNA, total RNA and viral RNA from a wide range of targets. The mix ensures high specificity and sensitivity in single and multiplex amplification, making it the best choice for extremely low-copy-number targets in pathogen detection.

4X CAPITAL™ qPCR Probe Master Mix Plus uses a proprietary enzyme and buffer formulation for efficient cDNA synthesis and qPCR in a single tube.

Handling

Prevention of contamination

RNase contamination is a concern when handling RNA. To minimize the risk of contamination and prevent RNA degradation, follow the guidelines below:

• Wear disposable gloves when handling the solutions.
• Dedicate separate sterile areas for the preparation of samples and reaction mixtures.
• Use molecular-grade nuclease-free water and reagents.
• Include a non-template control reaction in every PCR assay.

Basic Protocol

• Calculate the required volume for the desired number of reactions and include one additional reaction.
• Use high-quality PCR plates and seals designed for fluorescence applications.
• Reserve plate positions for positive controls (control RNA) and negative controls (water or buffer).
• For better correlation, run the reactions in triplicate.

• Mix and centrifuge briefly to collect the liquid at the bottom of the tube.
• Place the reaction into the PCR cycler.

Cycling program

*The recommended annealing/extension temperature is primer Tm +2 °C. Use gradient PCR to optimize the annealing temperature. Do not use annealing temperatures below 60 °C.

Product manual

Other documentation