5X One-Step Probe RT-qPCR Master Mix

Croyez 5X One-Step Probe RT-qPCR Master Mix is a one-step real-time reverse transcription-polymerase chain reaction (RT-qPCR) kit developed for cDNA synthesis and real-time PCR in the same tube. This product is suitable for probe-based detection and formulated as a 5-fold premix. Reaction can be simply set up by adding the RNA template, primers, and probes. This master mix does not contain ROX reference dye; it offers great convenience and minimizes the risk of cross-contamination.
No. Size Price Qty Status
C15012-1ML 1 mL $260.00 Inquiry
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The following procedure is a general guideline for One-step RT-qPCR reaction. To maintain an RNase-free environment, always wear disposable gloves, and use laboratory consumables and water of nuclease-free grade during the whole experiment course.

RT-qPCR reaction set-up:
1. Place all required reagents on ice.
Component Amount Final concentration
5X One-Step Probe RT-qPCR Master
4 μL 1X
Forward primer (10 μM) 0.8 μL 0.4 μL
Reverse primer (10 μM) 0.8 μL 0.4 μL
Probe (10 μM) 0.4 μL 0.2 μL
RNA template X μL ≦1 μg (total RNA)
Nuclease-Free H2O Y μL -
Total reaction volume 20 μL -
* See Usage Notes for additional guidelines on primer/template preparation.
2. Gently mix the reaction thoroughly to achieve uniform distribution and briefly centrifuge.
3. Thermal cycling conditions for standard qPCR
Step Cycles Temperature Time
Reverse transcription 1 50°C 10–15 min
Enzyme activation 1 95°C 5 min
Denaturation 40-45 95°C 5–15 sec
Annealing/Extension 55-65°C 30–60 sec