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Oncology & Haematology · EGFR
Biovendor Distributed by Goffin Diagnostics

Lung Cancer (EGFR)

The BioVendor fastGEN Lung Cancer kit prepares a sequencing library for EGFR by next-generation sequencing on an Illumina instrument. One kit covers 16 reactions, each with its own index. In non-small cell lung cancer the EGFR mutation status determines the choice of tyrosine kinase inhibitor. Exons 18 to 21 contain the activating deletions and substitutions as well as the resistance mutation T790M in exon 20.

RUO Biovendor 1 Unit (16 Reactions)
Brand
Biovendor
Packaging
1 Unit (16 Reactions)
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Product details

This kit in detail

  • EGFR exons 18, 19, 20 and 21
  • NGS library preparation in a single-step PCR, amplicons up to 170 bp
  • 16 reactions per kit, each with its own index
  • DNA from tumour tissue, including FFPE
  • About 4 hours in total, under 30 minutes hands-on
  • Illumina MiSeq or MiniSeq, analysis in GENOVESA

The BioVendor fastGEN Lung Cancer kit prepares sequencing libraries for genotyping EGFR by massively parallel sequencing. It is a library preparation kit, not a DNA extraction kit and not a real-time PCR assay: the DNA has to be isolated beforehand, and the result comes from the sequencer.

Regions covered

EGFR, exons 18, 19, 20 and 21. The manufacturer catalogue number is RDNGS0002.

Method

The kit uses a single-step PCR with hybrid primers that produce short amplicons of up to 170 bp. Sample and Master Mix are combined in about 10 minutes, the reaction runs on a real-time PCR instrument, and the pooled and purified libraries are sequenced on an Illumina instrument. Total workflow time is around 4 hours, of which less than 30 minutes is hands-on.

Sample and DNA input

The input is 5 µl of isolated DNA per reaction, at 10 to 200 ng/µl measured by spectrophotometer or from 1 ng/µl measured with the Qubit HS assay. DNA from tumour tissue, including formalin-fixed paraffin-embedded material, is used.

Kit contents

The kit contains 16 ready-to-use Master Mixes, each with its own index, plus the read 1 sequencing primer (R1SP), the read 2 sequencing primer (R2SP) and the index sequencing primer (ISP). The sequencing primers are added to positions 12, 13 and 14 of the Illumina cartridge alongside the Illumina primers. No positive or negative control is supplied. Storage is at -20 °C.

Analysis

Raw FASTQ data is analysed in the fastGEN module of the GENOVESA software, a cloud application supplied for this purpose.

  • Master Mix, 16 vials, each with a unique index
  • R1SP, read 1 sequencing primer
  • R2SP, read 2 sequencing primer
  • ISP, index sequencing primer

16 reactions per kit, REF RDNGS0002. No positive or negative control is supplied; the manufacturer recommends running controls of your own. DNA isolation reagents are not supplied. Storage at -20 °C until the expiry date. The sequencing primers are added to positions 12, 13 and 14 of the Illumina cartridge together with the Illumina primers.

Documentation

Datasheets and regulatory documentation

Everything your quality team needs for evaluation and procurement.

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