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FGFR RGQ RT-PCR Kit

For qualitative detection of alterations in the FGFR2 and FGFR3 genes
  • Detection of four point mutations and five fusions
  • High sensitivity and specificity
  • Simple workflow with next-day results
  • Automated data analysis using Rotor-Gene AssayManager v2.1 software

The FGFR RGQ RT-PCR Kit is a qualitative assay for the detection of four point mutations in of the FGFR3 gene: exon 7 [p.R248C (c.742C>T), p.S249C (c.746C>G)] and exon 10 [p.G370C (c.1108G>T), p.Y373C (c.1118A>G)]. In addition, the assay detects three fusions in the FGFR3 gene (FGFR3-TACC3v1, FGFR3-TACC3v3 and FGFR3-BAIAP2L1) and two fusions in the FGFR2 gene (FGFR2-BICC1 and FGFR2-CASP7). 

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Cat No./ID: 73504
RNeasy FFPE Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
Cat No./ID: 8747010
FGFR RGQ RT-PCR Kit (24)

For 24 reactions: Reverse Transcriptase, RT Buffer 1, RT Buffer 2, RT Primer Mix, Mutations-1 Reaction Mix, Mutations-2 Reaction Mix, Fusions-1 Reaction Mix, Fusions-2 Reaction Mix, Water for NTC, Water for sample dil., FGFR Positive Control, PC Diluent

Cat No./ID: 9001580
Rotor-Gene Q 5plex HRM Platform
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
Cat No./ID: 9001650
Rotor-Gene Q 5plex HRM System
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

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.


Product Details

Principle

The FGFR RGQ RT-PCR Kit is based on the selective amplification of alterations in the FGFR2 and FGFR3 genes using the Rotor-Gene Q 5plex HRM instrument for sensitive and specific analysis. The assay exploits the qPCR oligonucleotide hydrolysis principle using TaqMan probes.

Allele-specific technology allows accurate and highly reproducible detection of alterations, based on the use of specific forward and reverse primers and probes; only a perfect match between the primers and probes with the target cDNA allows extension and amplification in the PCR reaction. Data analysis is performed automatically. Outputs for each PCR are displayed individually, with "Amplification Detected" displayed when a specific FGFR mutation is detected.

Procedure

The fast and simple workflow takes ~12 hours from Sample to Insight.

RNA is first prepared from formalin-fixed paraffin-embedded (FFPE) urothelial tumor samples using the RNeasy DSP FFPE Kit or RNeasy FFPE Kit. Purified RNA is then reverse transcribed using Reverse Transcriptase to generate cDNA for real-time PCR analysis. Optimized reverse transcription reagents and PCR master mixes provided in the kit enable high-fidelity reverse transcription and sensitive real-time PCR on the Rotor-Gene Q 5plex HRM instrument. Qualitative results are displayed in Rotor-Gene AssayManager software, informing the system operator if one or more of the four point mutations and five fusions detected by the kit are present in each sample.

Applications

The FGFR RGQ RT-PCR Kit enables qualitative detection of four point mutations and five fusions in the FGFR2 and FGFR3 genes.

Product Resources

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Kit Handbooks (5)
FAQs (54)
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Quick-Start Protocols (2)
For purification of total RNA from formalin-fixed, paraffin-embedded tissue sections
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Instrument User Manuals (3)

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For use with Rotor-Gene Q Software version 2.3.4
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For Rotor-Gene Q instruments using Rotor-Gene Q Software version 2.3.4
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Operating Software (1)
For use on the Rotor-Gene Q. Rotor-Gene Q software 2.3.5 is compatible with Windows 7 and Windows 10 operating systems
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