RealMOD™ Probe M² 2X qRT-PCR mix
It is composed of TaqMan Probe type and is a 2X Master mix type product that can easily perform real-time RT-PCR (qRT-PCR) using RNA as a template.A TaqMan probe type product capable of one-step qRT-PCR using the extracted RNA as a template for cDNA synthesis and real-time PCR in a tube with RNA as a templateReal-time RT-PCR reaction volume control with 2X Master mix typeMinimize non-specific reactions by adding Anti Taq antibody based Hot-start DNA polymeraseReproducible experiments with excellent stability
RealMOD™ Probe M2 2X qRT-PCR mix is a TaqMan Probe type product capable of one-step qRT-PCR using extracted RNA as a template. Thermal stability and cDNA synthesis efficiency were improved by using genetically engineered M-MLV RTase for reverse transcription. In addition, non-specific reactions were minimized by adding Anti Taq antibody based Hot-start DNA polymerase. Reproducible experiments are possible by confirming excellent stability through cold thawing and accelerated aging tests. This product is provided in 2X Master mix type, so the capacity can be adjusted according to the experimenter.
| Content | Volumes |
|---|---|
| RealMOD™ Probe M2 2X qRT-PCR mix | 1 ml |
| Manual | 1 ea |
RealMOD™ Probe M2 2X qRT-PCR mix is a product capable of standard qRT-PCR. The types that can be tested using this product are as follows.
Applied BioSystems : QuantStudioTM 12K Flex, ViiATM 7, 7900HT, 7500, 7700, StepOneTM & StepOne plusTM
Bio-Rad : CFX96TM & CFX384TM, iQTM5 & MyiQTM, Chomo4TM, Opticon® 2 & MiniOpicon®
Qiagen : Rotor-Gene® Q, Rotor-Gene® 6000
Eppendorf : Mastercycler®, ep realplex2 & ep realplex4
Roche : LightCycler® 480
Illumina : The EcoTM
Figure 1. Amplification of SARS CoV-2 RNA using RealMOD™ Probe M² 2x qRT-PCR mix
SARS CoV-2 RNA was serial diluted 1/10, 10, 105, 104, 103, 102. Amplification of SARS CoV-2 RNA using RealMOD™ Probe M² 2X qRT-PCR mix on anABI 7500 fast Real-time PCR system. Real-time RT-PCR results; slope : -3.327, R2 : 0.999, PCR efficiency 99.8%
Figure 2. Performance comparison test with other products
Real-time PCR amplification of the RdRP gene of coronavirus. SARS CoV-2 RNA was serial diluted 1/10, 106, 105, 104, 103, 102. Amplification of SARS CoV-2 RNA using RealMOD™ Probe M² 2X qRT-PCR mix on an Bio-Rad CFX96 Real-time PCR system. Real-time RT-PCR results; M2 2X qRT-PCR has excellent ct value and dynamic range.
As a result of real-time PCR experiment, a dynamic range suitable for the dilution factor (1/10) was observed.
- It has been confirmed that the reliability of the standard curve is very high and the PCR efficiency is also excellent.
- Compared with other companies' products, excellent Ct value and stable PCR amplification curve were confirmed in the same sample.
Q. As a result of the experiment, amplification cannot be confirmed.
A. First, please check if there are any problems in the template extraction process. Secondly, when preparing PCR mixture, check that there are no missing contents (primer, probe, 2X qRT-PCR mix). Third, check the experimental conditions by reviewing the primer design and the annealing temperature of the prime.
Q. As a result of the experiment, a signal was detected in NC (negative control).
A. First, since non-specific reactions may occur due to environmental factors, please cleanly disinfect the equipment and experiment table you are using before proceeding. Secondly, since non-specific reactions may appear due to experimental factors (contamination by the template to be used), prepare a mixture, dispense negative control first, and conduct an experiment on the target.
Q. Is ROX dye added to the product?
A. ROX dye is not added to this product, and if necessary, it should be added afterwards. If there is a choice of passive reference dye in the experiment, proceed with none.
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