EZProbe One Step qRT-PCR Probe Kit
Type: Probe One Step qRT-PCR Kits
Catalog Number: EZB-RTProbe-S / EZB-RTProbe
Pack Size: 100 Rxns / 500 Rxns
Price: 525 USD / 2250 USD
Promotion: 420 USD / 1800 USD
Introduction
The EZBioscience® EZProbe One Step qRT-PCR Kit is a one-step kit to quantify RNA using fluorescent probe. With the exception of primers, probe and template, this kit contains in four vials all the components required to perform rapid, sensitive and reproducible qRT-PCR.
10× EZProbe One Step Enzyme Mix contains M-MLV (RNase H-) reverse transcriptase, RNase Inhibitor and Hot Start DNA Polymerase; 2× EZProbe One Step Buffer contains dNTPs and optimized buffer; 50× ROX Reference Dyes are used to rectify the error of fluorescence signals between different wells.
The M-MLV Reverse Transcriptase, which is an RNA-dependent DNA polymerase with a significantly attenuated RNase H activity, can synthesize cDNA at a temperature range of 42°C to 60°C and is inactivated during the activation step of the Hot Start DNA Polymerase.
RNase inhibitor can protect RNA templates from degradation.
This buffer has been optimized to allow both reverse transcription and PCR amplification to occur in the same reaction across a wide range of templates.
Hot Start DNA Polymerase requires an activation step at 95°C for 5 minutes, which prevents non-specific amplification during cDNA synthesis.
Components
Components | EZB-RTProbe-S (100 Rxns) | EZB-RTProbe (500 Rxns) |
10× EZProbe One Step Enzyme Mix*1 | 200 μl | 1 ml |
2× EZProbe One Step Buffer*2 | 1 ml | 5 ml |
50× ROX Reference Dye1*3 | 44 µl | 220 µl |
50× ROX Reference Dye2*3 | 44 µl | 220 µl |
*1: Contain reverse transcriptase, RNase Inhibitor, Hot Start DNA Polymerase.
*2: Contain dNTPs and optimized buffer.
*3: Used to rectify the error of fluorescence signals between different wells.
Storage
The kit should be stored at -20°C and protected from light. Avoid repeated freeze thawing. The quality of this product can be guaranteed for 18 months under recommended storage conditions.
Notice
Please select an appropriate ROX according to the Real-time PCR instrument used:
Do Not Use ROX | Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, MyiQ™, MiniOpticon™,Opticon®, Opticon 2, Chromo4™; Roche LightCycler™ 96, Roche LightCycler™ 480; Eppendorf Mastercycler® ep realplex, realplex 2s; Illumina Eco qPCR; Qiagen/Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene® 6000; Thermo Scientific PikoReal Cycler; Analytikjena qTOWER 3G; Cepheid SmartCycler®. |
Use ROX1 (1×) | ABI 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast; StepOne™, StepOne Plus™. |
Use ROX2 (1×) | ABI 7500, 7500 Fast, Quant-Studio 3, 5, 6, 7, 12K Flex, Dx, ViiA™7; Stratagene MX3000P™, MX3005P™, MX4000™. |
Representative Results
Figure1. GAPDH mRNA expression level in serially diluted RNA from SARS-CoV-2-pseudovirus were detected. The amplification plots above indicate that the target gene could be amplified efficiently and specifically.
For detailed information, please check the product manual.