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PIK3CA RGQ PCR Kit

For qualitative detection of 11 mutations in the PIK3CA gene by real-time PCR

Features

  • Reliable detection of relevant mutations in the PIK3CA gene
  • High sensitivity and specificity
  • Results in less than two working days
  • Optimized reagents and reaction mixes
  • Automated data analysis using Rotor-Gene AssayManager v2.1 software

Products

For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
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PIK3CA RGQ PCR Kit

Cat. No. / ID: 873101

For 24 reactions: 6 Reaction Mixes, Positive Control, Taq DNA Polymerase, Water for NTC, and Water for Sample Dilution
₹255,270.00
Add to cart
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Rotor-Gene Q 5plex HRM Platform

Cat. No. / ID: 9001580

Real-time PCR cycler and High Resolution Melt analyzer with 5 channels (green, yellow, orange, red, crimson) plus HRM channel, laptop computer, software, accessories: includes 1-year warranty on parts and labor, installation and training not included
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Rotor-Gene Q 5plex HRM System

Cat. No. / ID: 9001650

Real-time PCR cycler and High Resolution Melt analyzer with 5 channels (green, yellow, orange, red, crimson) plus HRM channel, laptop computer, software, accessories: includes 1-year warranty on parts and labor, installation and training
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QIAamp DNA FFPE Tissue Kit (50)

Cat. No. / ID: 56404

For 50 DNA preps: 50 QIAamp MinElute Columns, Proteinase K, Buffers, Collection Tubes (2 ml) Use our next-generation QIAamp DNA FFPE Advanced Kits for improved performance.
₹17,370.00
Add to cart
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QIAamp Circulating Nucleic Acid Kit (50)

Cat. No. / ID: 55114

For 50 preps: QIAamp Mini Columns, Tube Extenders (20 ml), QIAGEN Proteinase K, Carrier RNA, Buffers, VacConnectors, and Collection Tubes (1.5 ml and 2 ml)
₹108,510.00
Add to cart
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Rotor-Gene AssayManager v2.1

Cat. No. / ID: 9024203

Software for routine testing in combination with Rotor-Gene Q and Rotor-Gene Q MDx instruments. IMPORTANT: Does not include license for use. License key must be purchased separately. Visit the Download Page for download, installation and licensing instructions.

Product Details

The PIK3CA RGQ PCR Kit is a real-time qualitative PCR test for the detection of 11 mutations in the phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) gene using a sample of DNA extracted from either formalin-fixed, paraffin-embedded (FFPE) breast tumor tissue or K2EDTA anticoagulated blood plasma, taken from a patient with breast cancer.

 

Principle

The PIK3CA RGQ PCR Kit is comprised of  six reaction mixes; one control reaction targeting exon 15 and five mutation-specific reactions utilized to detect 11 mutations in exons 7, 9 and 20 of the PIK3CA gene (Exon 7: C420R; Exon 9: E542K; E545A, E545D [1635G>T only], E545G, E545K, Q546E, Q546R; and Exon 20: H1047L, H1047R, H1047Y). Allele-specific technology allows accurate and highly reproducible detection of mutations; DNA is selectively amplified using ARMS primers, probes and PCR clamps, with sensitive signal detection using the Rotor-Gene Q 5plex HRM instrument. Data analysis is performed automatically. Outputs for each PCR are displayed individually, with "Amplification Detected" displayed when a specific PIK3CA mutation is detected.

Procedure

The simple and straightforward testing workflow begins with manual DNA extraction from either FFPE breast tumor tissue (using the QIAamp DSP DNA FFPE Tissue Kit or QIAamp DNA FFPE Tissue Kit) or from K2EDTA anticoagulated plasma (using the QIAamp DSP Circulating Nucleic Acid Kit or QIAamp Circulating Nucleic Acid Kit), followed by sensitive real-time PCR on the Rotor-Gene Q 5plex HRM instrument. Rotor-Gene AssayManager software rapidly and accurately reports results, informing the system operator if one or more of the 11 mutations detected by the kit are present in each sample. The kit is a qualitative assay that can yield results in less than two working days.

Applications

The PIK3CA RGQ PCR Kit enables qualitative detection of 11 mutations in the PIK3CA gene

Resources

Brochures & Guides (7)
Sample to Insight solutions for successful molecular analysis
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Second edition — innovative tools
Now with even more applications!
Critical factors for molecular analysis of FFPE samples
Quick-Start Protocols (1)
Kit Handbooks (8)
For concentration and purification of free-circulating DNA and RNA from human plasma or serum
For verification of thermal accuracy of Rotor-Gene real-time cyclers
Q546R false mutation positive result
Scientific Posters (3)
Scientific Poster
PDF (671KB)
Fast and Integrated Screening Workflow to Assess Gene Expression of Individual Pathways
Safety Data Sheets (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
Instrument User Manuals (5)
For Rotor-Gene Q instruments using Rotor-Gene Q Software version 2.3.4
Guideline for ISO 20836:2020
For use with Rotor-Gene Q Software version 2.3.4
Operating Software (2)
To download Rotor-Gene AssayManager v2.1.1 after purchase:
    Click the Rotor-Gene AssayManager v2.1.1 Software header. On the next window, click Continue. Follow the on-screen instructions to install Rotor-Gene AssayManager v2.1.1.
    Rotor-Gene AssayManager v2.1.1 and license must be purchased prior to use.    

    IMPORTANT: In order to perform and analyze runs, at least one Rotor-Gene AssayManager v2.1.1 plug-in needs to be installed. Refer to your assay kit handbook for more information.

    IMPORTANT: To allow checksum integrity verification, use the value D24F10DEB7A1D60EE381E2AEA8789117D0980FCC.

To purchase a license key for Rotor-Gene AssayManager v2.1.1:
    1. After purchasing Rotor-Gene AssayManager v2.1.1 software and licenses, click the "Download" button and follow the onscreen instructions to install. After successful installation of the Rotor-Gene AssayManager v2.1.1 and a plug-in, a “key file” will be generated. This "key file" will be attached to the "Technical Support Form" in Step 5, below.   
    2. Go to the Technical Support Form. Make sure to include all required information and your Rotor-Gene Q or Rotor-Gene Q MDx serial number when filling out the form.       
    3. For Question 2, "Type of Request", select “Other”.
    4. For Question 4, “Your inquiry”, enter "Rotor-Gene AssayManager license request” and include your purchase order number, from your order confirmation.
    5. Include the "key file" from Step 1 as an attachment.
    6. Submit the form. QIAGEN Technical Services will respond via email with a license file within 1–2 working days.    
    7. After receiving your computer-specific Rotor-Gene AssayManager v2.1.1 license file, archive the license file locally on your computer.

IMPORTANT: The license file must always exist on the computer that has Rotor-Gene AssayManager v2.1.1 installed. The license file location should be protected and not subject to change.





For use on the Rotor-Gene Q. Rotor-Gene Q software 2.3.5 is compatible with Windows 7 and Windows 10 operating systems
Validation Reports (1)
Additional Resources (1)
Analysis Software (1)

FAQ

Will QuantiTect Primer Assays work with Rotor-Gene SYBR Green Kits using an annealing step at 60ºC?

Based on our extensive and successful testing of many QuantiTect Primer Assays with Rotor-Gene SYBR Green PCR Kits, we guarantee this.

 

 

FAQ ID -2124
If new dyes or chemistries appear on the real-time PCR market, will Rotor-Gene Q be capable of running those dyes?

The Rotor-Gene Q software allows creation of new excitation/detection wavelength combinations, which means that Rotor-Gene Q will work with dyes you may want to use in the future.

 

FAQ ID -2087
Do you have a protocol for Rotor-Gene software setup for the Rotor-Gene SYBR Green PCR and RT-PCR Kits?
Which downstream applications have been tested with SARS-CoV-2-derived RNA purified from saliva collected into PAXgene Saliva Collector?

RNA purified with the QIAamp Viral RNA Mini Kit has been used for quantification by qPCR with QuantiTect Probe RT-PCR Kit on QIAGEN Rotor-Gene Q.

3828
I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560
What is the composition of elution buffer ATE in the QIAamp DNA Investigator kit, QIAamp DNA FFPE Tissue kit and the QIAamp Fast DNA Stool Mini kit?

The composition of Buffer ATE is:

- 10 mM Tris-Cl pH 8.3

- 0.1 mM EDTA

- 0.04% NaN3 (sodium-azide)

FAQ ID -3122
How is the hypothesis test performed in the REST software?
In the REST 2009 software, the hypothesis test performs 10,000 random reallocations of samples and controls between the groups, and counts the number of times the relative expression on the randomly assigned group is greater than the sample data.
FAQ ID -2459
How do Rotor-Gene Probe Kits compare with QuantiFast Probe Kits?

Rotor-Gene Probe Kits are specially developed for Rotor-Gene cyclers. The unique rotary system of the cyclers combined with the kits’ proprietary buffer system enable ultrafast cycling. Rotor-Gene Probe Kits do not contain ROX dye.

 

FAQ ID -2125
Can 0.1 ml and 0.2 ml tubes used on the Rotor-Gene Q be labeled?

Both types of tubes run on the Rotor-Gene Q, and can be labeled on the top without any interference with data acquisition, because signal is detected from the bottom of the tube. Labeling is helpful and can prevent possible sample mix up.

 

FAQ ID -2082
Does the Rotor-Gene Q software have the capacity to export data?

There are several features available to allow exporting data across platforms to a variety of software packages. Sample names can easily be imported into the Rotor-Gene Q Excel sample sheets. By using a function "Export to Excel" the software automatically exports any results table in the software to Excel. Furthermore, reports can now be exported as Word files, which make the data compatible with any Microsoft office/Mac application. Reports can also be saved or sent in HTML format. All figures and graphs can be exported as Jpeg or Bitmap files for use in any desktop publishing software.

 

FAQ ID -2089
Do the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment?

No. The master mixes in Rotor-Gene Kits contain dTTP instead of dUTP. If UNG treatment is required, we recommend using QuantiTect +UNG Kits. QuantiTect Kits are also compatible with the Rotor-Gene Q; however, the kits require a significantly longer cycling time.

 

 

FAQ ID -2117
In the REST software, what is the range indicated by 'Std. Error' on the 'Results' page?
In the REST 2009 software, the range for the Standard Error on the results tab is 68% C.I. (confidence interval).
FAQ ID -2455
Is a passive internal reference dye, like ROX, required on the Rotor-Gene Q to obtain reproducible results?

No. The fixed optical path ensures uniform illumination and detection from sample to sample eliminating the need to use a reference dye such as ROX on the Rotor-Gene Q.

 

 

FAQ ID -2079
Is it possible to edit the sample sheet in the Rotor-Gene Q software after a PCR run?

Yes. The sample sheet can be amended during or after a PCR run by activating the 'Edit samples' button in the Rotor-Gene Q software. The sample sheet will open and may be modified.

 

FAQ ID -2092
Can data be analyzed on the Rotor-Gene Q while a run is in progress?

Yes, data can be analyzed on the Rotor-Gene Q while a run is in progress, allowing to save time for planning and setting up new experiments. Multiple copies of the Rotor-Gene Q software can be opened simultaneously during a run allowing analysis of previous experiments while the system is running.

 

FAQ ID -2091
What is a QuantiTect Primer Assay?

QuantiTect Primer Assays are primer pairs designed and bioinformatically validated specifically for real-time RT-PCR with SYBR Green detection. To find a primer assay for your target gene of interest, please visit our GeneGlobe data base.

For best results, we strongly recommend using QuantiTect Primer Assays in combination with QIAGEN's products for SYBR Green-based Real-Time PCR and RT-PCR.

FAQ ID -1141
If Vapor-Lock is used to overlay PCR reactions, which volume should be entered as reaction volume in the Rotor-Gene Q Software?

The reaction volume to be entered in the Rotor-Gene Q software should be the sum of the PCR reaction volume and the volume of Vapor-Lock.

 

 

FAQ ID -2150
How many signal readings are taken during the real-time data acquisition on the Rotor-Gene Q?

At the selected data acquisition step of the thermal cycle on the Rotor-Gene Q, the fluorescent signal intensity is measured 20 times for each sample as they spin past the detector. Tubes on a rotor spin past the excitation/detection optics every 150 milliseconds. The average of the 20 readings is then taken as the fluorescence of the sample at that cycle number.

 

FAQ ID -2083
Why is a 2-step (and not a 3-step) cycling protocol recommended for Rotor-Gene SYBR Green Kits?

This type of cycling allows a significant reduction in cycling time for Rotor-Gene SYBR Green PCR Kits. It is more effective than reducing the individual times for annealing and extension.

 

FAQ ID -2122
What types of reaction vessels are required for use in the Rotor-Gene Q?

We strongly recommend use of the dedicated Rotor-Gene Q Accessories, e.g., PCR Tubes 0.2 ml or Strip Tubes and Caps 0.1 ml provided by QIAGEN. These tubes are designed to give low background and perfectly match the Rotor-Gene Q requirements.

 

FAQ ID -2085
Why is the initial activation step different for Rotor-Gene Probe, SYBR Green and Multiplex Kits?

The buffer composition, which affects the initial reactivation of HotStarTaq Plus DNA Polymerase, has been optimized for each respective Rotor-Gene Kit.

 

FAQ ID -2118
How long does an Investigator Quantiplex run take?
On the Rotor-Gene Q, an Investigator Quantiplex run of 40 cycles takes approximately 48 minutes. On the Applied Biosystems 7500 Real-Time PCR System, an Investigator Quantiplex run of 40 cycles takes approximately 57 minutes
FAQ ID -2566
How many samples can be run in parallel with the Rotor-Gene Q real-time PCR instrument?

The Rotor-Gene Q instrument can be used with two different rotor formats: using tubes or rotor-dics. Tubes can be run in 36 and 72-well rotors and rotor-discs in Rotor-Disc 72 and Rotor-Disc 100 rotors. When programming the temperature profile please make sure the correct rotor type is selected.  

 

 

FAQ ID -1519
Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with Rotor-Gene Probe Kits?

Yes, use the assays at a final concentration of 1x with Rotor-Gene Probe Kits on the Rotor-Gene Q cycler.

See trademarks.  

FAQ ID -2126
How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling?

Our unique multiplex PCR buffer system with ammonium and potassium ions and Factor MP has been further optimized in QuantiFast and Rotor-Gene Kits. We have also discovered Q-Bond, a buffer component which supports the rapid formation of the polymerase–primer–template complex, leading to reduced annealing times.

FAQ ID -1430
Which qPCR instrument should I use with your RT² qPCR Primer Assays?

Our RT² qPCR Primer Assays may be used on any real-time instrument. qPCR solutions are available for the most popular qPCR instrumentation, including those from QIAGEN, ABI, BioRad, Stratagene.

Instrument-specific protocols are available for selected instruments, and can be accessed at the following link: http://www.sabiosciences.com/pcrarrayprotocolfiles.php

FAQ ID -2714
For which real-time cyclers has the Investigator Quantiplex Kit been validated?
The Investigator Quantiplex Kit has been validated for the Rotor-Gene Q, and the Applied Biosystems 7500 Fast and 7500 Real-Time PCR Systems.
FAQ ID -2567
How is the log graph threshold value (Ct) for the Rotor-Gene Q determined?

Based on the defined standards present, the automatic threshold function of the Rotor-Gene Q scans through all the possible threshold levels until the best fit is determined. This is defined as the R value or correlation coefficient, which is maximized to most closely approach 1.0. If there is no standard present, the threshold can also be set manually.

 

FAQ ID -2088
Which version of the Rotor-Gene Q software can be used to run the Investigator Quantiplex?
Using the Investigator Quantiplex on the Rotor-Gene Q: All versions of the Rotor-Gene Q software can be used to run the Investigator Quantiplex.
FAQ ID -2568
Do limiting primer concentrations need to be determined when using the Rotor-Gene Multiplex PCR Kit?

No, that is not necessary. Simply use the primer concentrations specified in the protocols in the handbook supplied with your Rotor-Gene Multiplex PCR Kit.

 

 

FAQ ID -2128
What real-time cycler should I use for my qPCR experiments?
There are several manufacturers of high-quality real-time cyclers. These include QIAGEN's Rotor-Gene Q, Applied Biosystems, BioRad, Stratagene, Eppendorf, Roche, TaKaRa, Fluidigm, and Cepheid. The important thing to keep in mind is that, once you select an instrument to use, you must use compatible Rotor-Gene Discs and tubes, 96 or 384 well plates, and qPCR master mixes that are optimized for use in that particular instrument.  For example, QIAGEN's Rotor-Gene SYBR Green, Probe, and Multiplex real-time master mixes.  
FAQ ID -2670
Is it possible to import a standard curve from a previous PCR run on the Rotor-Gene Q?

Yes. If the reaction efficiency between two PCR runs is not expected to vary, importing a standard curve from a previous run allows to estimate concentrations when a standard curve for the current run is not available. Curves can be imported from another channel, or from another run by clicking on 'Import Curve' in the Rotor-Gene Q software. Standard curves can be adjusted such that only the efficiency of the source curve is imported into the current run. Whether a standard curve should be adjusted depends on the PCR chemistry used. To adjust a standard curve, use a reference with a known concentration in the target run.

 

FAQ ID -2093
Are Rotor-Gene Kits compatible with reaction setup using the QIAgility instrument?

The majority of the Rotor-Gene Kit data shown in our literature has been generated with the help of the QIAgility instrument. We did not observe any problems during the pipetting steps.

 

FAQ ID -2116
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with the Rotor-Gene Multiplex PCR Kit?

Yes, simply use the assays at a final concentration of 1x with the Rotor-Gene Multiplex PCR Kit.

See trademarks

FAQ ID -2131
Can Vapor-Lock be used with samples processed on the QIAgility?

Vapor-Lock is fully compatible with the QIAgility instrument for high-precision, automated reaction setup. It is also highly suited for use with the Rotor-Gene Q cycler. For support to program your QIAgility instrument for use with Vapor-Lock, please contact your local QIAGEN Technical Support Department.

 

 

FAQ ID -2153
Do QuantiTect Primer Assays also work with Rotor-Gene SYBR Green PCR Kits?

We have performed numerous tests comparing the performance of Rotor-Gene SYBR Green PCR Kits and QuantiTect Kits with QuantiTect Primer Assays. Due to the optimized ion concentrations in the PCR buffers, both perform equally well with QuantiTect Primer Assays and do not require any adjustment of primer concentration.

 

FAQ ID -2123
During data analysis, how should the threshold be set in the Yellow channel on the Rotor-Gene Q cycler to analyze the Internal Control from the QuantiFast Pathogen PCR +IC Kit or the QuantiFast Pathogen RT-PCR +IC Kit
On the Rotor-Gene Q, setting the threshold in the Yellow channel to an absolute value of 0.05 will give satisfactory results in most cases.
FAQ ID -2607
What is the average amount of DNA and RNA present in 1 ml normal serum?

According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.

For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.

FAQ ID -635
What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits?

Rotor-Gene Kits are specifically developed for the Rotor-Gene Q PCR Cycler. The unique rotary system of the cycler combined with the kits’ proprietary buffer system enable ultrafast cycling. Rotor-Gene Kits do not contain ROX dye since no normalization to a passive reference is required. Also, Rotor-Gene Kits do not contain dUTP; therefore, UNG pretreatment is not possible.

 

FAQ ID -2119
Does the centrifugal force have any effect on the kinetics of a PCR reaction on the Rotor-Gene Q?

No. The speed at which the samples spin on the Rotor-Gene Q is not high enough to apply any significant centrifugal force on them.

 

 

 

FAQ ID -2081
How fast does the rotor spin during a run on the Rotor-Gene Q?

The sample rotor of the Rotor-Gene Q spins continuously at a speed of 400 rpm during a run.

 

FAQ ID -2080
Can the Rotor-Gene Multiplex RT-PCR Kit be used on other cyclers?

The specific features of Rotor-Gene Kits and Rotor-Gene cyclers work synergetically to enable an ultrafast-cycling protocol. We do not guarantee that the performance of the Rotor-Gene Multiplex RT-PCR Kit with the same cycling protocol will be the same on other cyclers.

 

 

FAQ ID -2142
Will Rotor-Gene Kits also work on the Rotor-Gene 6000 and 3000 cyclers?

Yes. Rotor-Gene Kits will also work on the Rotor-Gene 6000 and Rotor-Gene 3000 PCR cyclers with the cycling conditions specified in the Rotor-Gene kit handbooks.

 

FAQ ID -2121