qPCRBIO SyGreen 1-Step Go Hi-ROX

Art No | PB25.32

qPCRBIO SyGreen 1-Step Go Hi-ROX

Choose size / variation:

from 120.00 CHF

qPCRBIO SyGreen 1-Step Kits have been designed for fast, highly specific and ultra-sensitive cDNA synthesis and real-time PCR in a single tube.

Description

qPCRBIO SyGreen 1-Step Kits have been designed for fast, highly specific and ultra-sensitive 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.

Select from qPCRBIO SyGreen 1-Step Detect for the detection of extremely low copy number targets, or qPCRBIO SyGreen 1-Step Go for rapid and accurate results from high RNA template concentrations.

Thermostable reverse transcriptase 45°C to 55°C
Advanced RNase inhibitor
Non-PCR inhibiting intercalating dye
Rapid extension rate for early Ct values
Market-leading sensitivity
Increased limit of detection
Antibody-mediated hot start PCR
Compatible on all standard and fast cycling real-time PCR platforms

qPCRBIO SyGreen 1-Step Kits can be used to quantify any RNA template including mRNA, total RNA and viral RNA sequences. qPCRBIO SyGreen 1-Step Detect is designed for sensitivity and is ideally suited to the detection of extremely low copy number targets. qPCRBIO SyGreen 1-Step Go gives the earliest Ct and is formulated for rapid and accurate results from high template concentrations.

The kits include a thermostable and extremely active modified MMLV reverse transcriptase and advanced RNase inhibitor that prevents degradation of RNA by contaminating RNase. PCR amplification is powered by antibody-mediated hot start technology, preventing the formation of primer dimers and non-specific products leading to improved reaction sensitivity and specificity. Combining the latest developments in polymerase technology and advanced buffer chemistry we offer market- leading performance with minimal or no optimisation.

Absolute quantification
Relative gene expression analysis
Detection of extremely low copy number targets
qPCRBIO SyGreen 1-Step Detect recommended for template amounts of 1pg - 10ng total RNA or >0.01pg mRNA per reaction
qPCRBIO SyGreen 1-Step Go recommended for template amounts of 10pg - 100ng total RNA or >0.01pg mRNA per reaction

Use our qPCR Selection Tool to find out which ROX variant is compatible with your instrument.

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.

Material Safety Data Sheet (PDF)

Product manual (PDF)

Instrument Compatibility Tool

FAQs

qPCRBIO SyGreen 1-Step Detect | 1-Step Go

Figure 1.

Comparison of qPCRBIO SyGreen 1-Step Go
(purple) against competitor Bioline (blue)

Shows amplification traces of the ACTG1 gene from a dilution series of total RNA extracted from mouse liver. Total RNA concentration varied from 25pg to 250ng per 20μl reaction. Cycling conditions were 45°C 10 minutes for cDNA synthesis, followed by 95°C 2 minutes hot start, then 45 cycles of 95°C 10sec, 60°C 10sec on Roche LC480. qPCRBIO SyGreen 1-Step Go had equal performance at high RNA concentrations and superior performance at lower RNA concentrations, displaying linear spacing between amplification curves, earlier amplification by 3–4 cycles, and lower prevalence of primer dimer.

Figure 2 a.|b.

Comparison of qPCRBIO SyGreen 1-Step Detect (blue) and qPCRBIO SyGreen 1-Step Go (purple), showing ideal template ranges for each product.

The ACTG1 gene was amplified from a dilution series of total RNA extracted from mouse liver. qPCRBIO SyGreen 1-Step Go (purple) shows efficient amplification of total RNA in the range 100pg to 100ng per reaction. qPCRBIO SyGreen 1-Step Detect (blue) shows efficient amplification of total RNA in the range 10pg to 10ng per reaction (figure 2a). Cycling conditions were 45°C 10min for cDNA synthesis, followed by 95°C 2min
hot start, then 45 cycles of 95°C 10sec, 60°C 30sec, concluding with a melt analysis (figure 2b) on a Roche LC480.

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: