Probe-based RT-qPCR

Product description This product is a closed-tube fluorescent quantitative RT-qPCR system based on RNA templates for simultaneous detection of multiple targets. It integrates the reverse transcription and quantitative amplification processes, significantly simplifying operational steps and maximizing control of cross-contamination risks. The kit features two core enzyme technologies: a high-performance thermostable reverse transcriptase for efficient first-strand cDNA synthesis, and a high-performance Taq DNA polymerase modified for hot-start to ensure precise quantitative amplification. The premixed system includes a specially optimized multiplex reaction buffer system that ensures independent amplification efficiency for each primer pair while enabling parallel detection of multiple targets. This product is designed for research needs such as multiplex pathogen detection, gene expression profiling, and simultaneous quantitative analysis of multiple indicators in complex samples. Specifications Catalog Number N132052E N132052S N132052M Specifications 50 T 100 T 1000 T Components Component Identification Component Name N132041E N132041S N132041M N132052-A 2×RT-qPCR Buffer 750 μL 1.5 mL 15 mL N132052-B 2×RT-qPCR Enzyme Mix 60 μL 120 μL 1.2 mL 【Notes】 1) The Buffer mainly contains dNTPs, dUTP, Mg², stabilizers, etc. 2) The Enzyme Mix mainly contains a high-performance thermostable reverse transcriptase, Taq enzyme, RNase inhibitor, and UDG enzyme. Storage Store at -25 to -15℃ in the dark. Valid for 18 months. Notes 1. Please prepare the reaction mixture inside a laminar flow cabinet and use nuclease-free pipette tips and reaction tubes. Filter-tipped pipette tips are recommended to avoid cross-contamination and aerosol contamination. 2. For your safety and health, please wear a lab coat and disposable gloves when operating. 3. For Research Use Only. Instructions 1. Recommended Reaction System Component Volume（μL） Final concentration 2×RT-qPCR Buffer 15 1× 2×RT-qPCR Enzyme Mix 1.2 - Primer/Probe mix (2.5 μM) 3 0.25 μM Template RNA 1-10 - RNase Free H2O Up to 30 - 【Notes】 1) Ensure thorough mixing before use to avoid excessive bubbles from vigorous shaking. 2) The Primer/Probe Mix contains multiple pairs of primers and probes. The final concentration of each primer can be adjusted between 0.1-1.0 μM depending on the situation. The final concentration of the probes can also be adjusted within the range of 0.05-0.5 μM based on the requirements for the fluorescence curve and signal peak. 3) Due to the high sensitivity of qPCR, it is recommended to dilute the template. Aim to keep the Ct values between 20 and 35 for optimal results. 2.  Reference Amplification Program Recycling procedure Temperature Time Cycle Number Reverse transcription 50℃ 20 min 1 Reverse transcription 95℃ 5 min 1 Amplification reaction 95℃ 15 sec 40-45 60℃ 30 sec 【Notes】 1) Reverse transcription can be performed at either 42℃ or 55℃. 2) The amplification reaction temperature should be adjusted according to the Tm value of the designed primers. 3) The required fluorescence signal acquisition time varies among different qPCR instruments. Please set it according to the shortest time limit. 3. Compatible Instruments Applied Biosystems: 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™ , StepOne Plus™ , 7500, 7500 Fast, ViiA™ 7, QuantStudio™ 3 and 5, QuantStudio™ 6,7,12k Flex; Bio-Rad: CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4; Eppendorf: Mastercycler ep realplex, realplex 2 s; Qiagen: Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000; Roche Applied Science: LightCycler 480, LightCycler 2.0; Lightcycler 96; Stratagene: MX3000P™, MX3005P™, MX4000P™; Thermo Scientific: PikoReal Cycler; Cepheid: SmartCycler; Illumina: Eco qPCR; SLAN: SLAN-96S,SLAN-96P.  

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