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Rotor-Gene SYBR® Green PCR Demo Kit

For evaluation of Rotor-Gene Q performance

Features

  • Evaluate Rotor-Gene Q performance with Rotor-Gene master mixes
  • Determine the accuracy and reproducibility of the Rotor-Gene Q
  • A complete system containing all reaction components
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Rotor-Gene SYBR Green PCR Demo Kit (80)

Cat. No. / ID: 204001

For 80 reactions: 2x Rotor-Gene SYBR Green PCR Master Mix, 10x QuantiTect Primer Assay for GPER, Standards, Unknown Samples, Buffer TE, RNase-Free Water
The Rotor-Gene SYBR® Green PCR Demo Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The Rotor-Gene SYBR Green PCR Demo Kit is for use in demonstrating the performance of the Rotor-Gene Q in combination with Rotor-Gene master mixes. The kit contains all the necessary reaction components to quantify a genomic DNA target by SYBR Green-based real-time PCR. These include prediluted genomic DNA standards, 2 unknown samples, an optimized master mix, and a primer pair specific for the human G protein-coupled estrogen receptor 1 (GPER) gene. Using the kit, the reliability, reproducibility, and sensitivity of gene quantification with the Rotor-Gene Q can be evaluated.

Principle

High-precision in real-time PCR can be achieved using the combination of Rotor-Gene Kits and the Rotor-Gene Q real-time PCR cycler. The ready-to-use master mix supplied with Rotor-Gene Kits allows fast and accurate gene quantification without the need for reaction optimization, while the Rotor-Gene Q employs a unique centrifugal rotary design. PCR tubes are placed into a rotor that spins tubes past a single excitation light source and a single detector in a chamber of moving air. This means that there is minimal optical and temperature variation between tubes, enabling high precision in real-time PCR quantification. In addition, as the rotor spins continuously at 400 rpm, high-speed data acquisition is possible.

With the Rotor-Gene SYBR Green PCR Demo Kit, highly specific amplification is assured through a balanced combination of ions that minimizes nonspecific primer annealing (see figure "  Specific primer annealing"). Fast cycling without compromising performance is achieved using Q-Bond, a novel PCR additive that enables cycler run times of as low as 45 minutes (see figure "  Fast primer annealing").

See figures

Procedure

Using the Rotor-Gene SYBR Green PCR Demo Kit, SYBR Green-based real-time PCR is carried out to quantify different copy numbers of a genomic DNA target. Each reaction consists of:

  • Human genomic DNA template of a defined copy number
  • Rotor-Gene SYBR Green PCR Master Mix
  • QuantiTect Primer Assay specific for the human G protein-coupled estrogen receptor 1 (GPER) gene

The Rotor-Gene SYBR Green PCR Demo Kit handbook contains 3 protocols. Rotor-Gene users follow either the protocol for manual reaction setup or the protocol for automated reaction setup using the QIAgility. After reaction setup, users then proceed to the protocol for real-time PCR on the Rotor-Gene Q.

Manual pipetting steps can be avoided by using the QIAgility, a compact benchtop instrument that provides rapid, high-precision PCR setup. Mistakes in reaction setup due to human error are reduced and may be eliminated. The QIAgility perfectly complements the combination of the Rotor-Gene Q and Rotor-Gene Kits, enabling easy dispensing of liquids into tubes, strip tubes, and Rotor-Discs.

A standard curve is generated from the CT values obtained from a set of standards (2000, 1000, 500, 250, and 125 copies; each standard is analyzed in quadruplicate). The standard curve is then used to determine the copy number for 2 unknown samples (500 and 250 copies; 24 replicates of each unknown sample are analyzed [when performing manual reaction setup, a minimum of 4 replicates for each unknown sample can be set up instead]). In addition, 4 no template control (NTC) reactions are carried out. Thus, a total of 72 reactions are run at the same time on the Rotor-Gene Q.

Applications

The Rotor-Gene SYBR Green PCR Demo Kit is for use in demonstrating the performance of Rotor-Gene Kits in combination with the Rotor-Gene Q. The kit is also compatible with the Rotor-Gene 3000 or the Rotor-Gene 6000. Using this kit, the accuracy and reproducibility of gene quantification with the Rotor-Gene Q can be evaluated.

Supporting data and figures

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (1)
For evaluation of the performance of the Rotor-Gene Q

FAQ

How do I create a workspace that is free of DNA contamination, prior to carrying out a qPCR experiment?

Any DNA contamination will artificially inflate the SYBR Green signal, yielding skewed gene expression profiles and false-positive signals. The most common source of DNA contamination is from PCR products generated during previous experiments. Such contamination is most often due to the improper disposal of tubes, tips, and gels that previously came into contact with PCR products. Additionally, PCR products may also contaminate pipettors, racks, work pads, and commonly used reagents such as water and buffers. To minimize the risk of contaminating your experiment with extraneous DNA, the following steps should be taken:

 

  • Remove a single aliquot of water from your PCR-grade stock, sufficient to complete the experiment. This minimizes the number of times that the stock container is opened, thereby minimizing contamination risks.
  • Use only fresh PCR-grade reagents and disposable labware.
  • Treat any labware (tubes, tips, and tip boxes) used in PCR with 10% bleach, before discarding.
  • Maintain a dedicated workspace for PCR setup (perhaps a PCR-only hood), away from areas of the lab where post-PCR work is done, such as running gels, enzyme digestions, and cloning.
  • Change the lab bench pads/papers often and decontaminate lab benches and labware (racks, pipettors, etc.) before each use by washing with 10% bleach, and/or exposing to UV light for at least 10 minutes. This serves to degrade and/or inactivate contaminating DNA.
  • Before, during, and after the experiment, minimize the opening and closing of any tubes or plates used during the experiment.  
FAQ ID -2654