qPCRBIO Probe 1-Step Go Lo-ROX 1x500ml | 50000x20μl reactions – Witec AG

Art No | PB25.41-500
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Size: 1x500ml | 50000x20μl reactions

qPCRBIO Probe 1-Step Go is a universal probe kit designed for fast, highly specific and ultra-sensitive probe-based real-time RT-PCR.

Description

The kit is engineered for use on a wide range of probe technologies including TaqMan®, Scorpions® and molecular beacon probes.Rapid and sensitive detection of RNA viruses including SARS-CoV-2.

  • Rapid extension rate for early Ct values
  • Market-leading sensitivity – increased limit of detection
  • High efficiency in multiplex reactions
  • Advanced RNase inhibitor
  • Antibody-mediated hot start PCR
  • Compatible with all real-time PCR platforms – standard and fast cycling conditions

qPCRBIO Probe 1-Step Go is a universal probe kit designed for fast, highly specific and sensitive probe-based real-time RT-PCR. We use the latest developments in reverse transcriptase technology and buffer chemistry to give efficient cDNA synthesis and real-time PCR in a single tube.

qPCRBIO Probe 1-Step Go is engineered for use on a wide range of probe technologies including TaqMan®, Scorpions® and molecular beacons probes. The kit includes a 2x mix and can be used to quantify any RNA template including mRNA, total RNA and viral RNA sequences. qPCRBIO Probe 1-Step Go is designed to give rapid and accurate results over a broad range of template concentrations and is ideally suited to the detection of RNA viruses including SARS-CoV-2.

The kit includes a thermostable and extremely active modified MMLV reverse transcriptase (RTase Go) and advanced RNase inhibitor that prevents degradation of RNA by contaminating RNase. Antibody-mediated hot start technology prevents the formation of primer dimers and non-specific products giving highly specific and ultra-sensitive real-time RT-PCR with unrivalled efficiency in multiplex. Combining the latest developments in polymerase technology and advanced buffer chemistry we offer market-leading performance with minimal or no optimisation.

 

  • COVID-19 / SARS-CoV-2 testing and research
  • Diagnostic real-time PCR
  • Multiplex or singleplex
  • Detection of low copy number targets
  • Absolute quantification
  • Relative gene expression analysis
  • TaqMan®, Scorpions® and molecular beacon probes

Storage

The kit should be stored between -30°C and -15°C. Avoid prolonged exposure to light. If stored correctly the kit will retain full activity for 12 months. The kit can be stored at 4°C for 1 month. The kit can go through 30 freeze/thaw cycles with no loss of activity.

qPCRBIO Probe 1-Step Go

Figure 1.

High efficiency and broad dynamic range

Shows TaqMan® probe amplification traces of mouse gene ACTB using mouse liver total RNA as template in triplicate. Template concentrations are 10x serial dilutions ranging from 10pg to 1µg total RNA per 20µl reaction. Cycling conditions were 45°C 10min, 95°C 3min, then 45 cycles of 95°C 10s, 60°C 30s. qPCRBIO Probe 1-Step Go shows high efficiency over a broad dynamic range.

Figure 2. qPCRBIO Probe 1-Step Go in multiplex

Figure 2.

qPCRBIO Probe 1-Step Go in multiplex

Four mouse housekeeping genes were amplified simultaneously in a single multiplex reaction. 1μg
of mouse liver total RNA was used as template. Amplification was detected using TaqMan probes in the following gene/probe combinations: B2MG/HEX, ACTB/Cy5, GAPDH/FAM, and ACTG/TexasRed. Cycling conditions were 45°C 10min, 95°C 3min, then 45 cycles of 95°C 10s, 60°C 30s. This demonstrates that the qPCRBIO Probe 1-Step Go mix can be used to quantify and compare expression levels of multiple genes in a single reaction.

FAQs ROX

ROX (6-carboxy-X-rhodamine) is used as a passive reference dye in ROX-dependent real-time PCR instruments to normalize for variations of fluorescence levels that can arise mainly due to optical path variations among wells. Normalisation of the fluorescence intensity (Rn) is done in real-time PCR software by dividing the emission intensity of the specific signal by the emission intensity of ROX.

ROX does not take part in the PCR reaction and its fluorescence levels are not proportional to the quantity of DNA in each well, so the addition of this fluorophore to a mix provides a constant fluorescent signal during amplification.

Different types of real-time PCR instruments requiring a passive reference standard have different optimal concentrations of ROX, mainly due to the different optical configurations of each system (i.e. the different type of excitation source and optics used).

The addition of either too little or too much ROX would result in a very noisy signal impacting on the results of the reaction. Therefore, it is extremely important for the user to:

  • Determine the correct ROX concentration to optimise real-time PCR results, and
  • Check the ROX settings on the software used to set up the reaction

ROX concentrations in qPCRBIO Probe 1-Step Go Mixes:

The qPCRBIO Probe 1-Step Go Mixes come in 3 different formulations, each with a different concentration of ROX:

  • qPCRBIO Probe 1-Step Go Lo-ROX (PB25.41) contains 112 nM ROX
  • qPCRBIO Probe 1-Step Go Hi-ROX (PB25.42) contains 1.12 µM ROX
  • qPCRBIO Probe 1-Step Go Separate ROX (PB25.44) contains no ROX and include a separate tube of 50 µM ROX additive. This enables you to choose what concentration of ROX you’d like to use.

Available sizes:

PB25.41-01 100 x 20μL Reactions
PB25.41-03 300 x 20μL Reactions
PB25.41-05 500 x 20μL Reactions
PB25.41-12 1200 x 20μL Reactions
PB25.41-50 5000 x 20μL Reactions (1 x 50mL)
PB25.41-500 50’000 x 20μL Reactions (1 x 500mL)

Supplier

PCR Biosystems
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Specifications

2x qPCRBIO Probe 1-Step Go Lo-ROX

1 x 500mL

20x RTase Go with RNase Inhibitor

1 x 50mL

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