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2X One-Step Probe RT-qPCR Master Mix

Croyez 2X 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 2-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
C15011-1ML 1 mL $128.00 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
2X One-Step Probe RT-qPCR Master Mix 10 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