Rapid one-step RT-qPCR using PrimeDirect Probe RT-qPCR Mix

PrimeDirect Probe RT-qPCR Mix is designed for one-step, real-time RT-PCR via probe detection (5′-nuclease method). This product is supplied as a 2X premix to facilitate the easy preparation of reaction mixtures. It requires only the addition of your primer, probe, and sample, and the RT-qPCR can be performed by simply adding your sample directly to the reaction mixture without intervening nucleic acid purification steps.

The protocol allows all steps, from extraction of the nucleic acid to reverse transcription and qPCR, to be performed in a single tube, greatly reducing hands-on time and risk of contamination. The single-tube workflow also allows the detection of RNA viruses with complex higher-order structures.

High-efficiency PCR after cDNA synthesis detects DNA amplification in real time by measuring increased fluorescence from the probe. Because this product is highly resistant to various PCR inhibitors such as heparin (blood) and humic acid (soil), it can be used to directly detect pathogens, including viruses and bacteria, in various biological specimens and to directly analyze gene expression in cells.

Overview

• One-step RT-qPCR premix for ease of use
• Strong thermostability
• High tolerance for PCR inhibitors (e.g., heparin from blood and humic acid from soil)
• Compatible with direct addition of crude extracted RNA or samples to reaction mix
• Suitable for all probe-based assays

Applications

• Gene expression
• Pathogen detection (viruses and bacteria)
• Direct RT-qPCR with no intervening purification steps