Solid Cancer Kit (KRAS, NRAS, BRAF)
The BioVendor fastGEN Solid Cancer kit prepares a sequencing library for KRAS, NRAS and BRAF by next-generation sequencing on an Illumina instrument. One kit covers 16 reactions, each with its own index. The mutation status of KRAS, NRAS and BRAF determines treatment in colorectal carcinoma and in other solid tumours: a mutation in KRAS or NRAS predicts a lack of response to anti-EGFR antibodies, and BRAF V600E marks a subgroup with its own prognosis and treatment.
Quote-based, no public pricing. Add to your quote and our specialists put together a proposal with the right documentation, typically within 24 hours.
This kit in detail
- KRAS and NRAS hotspot codons and BRAF codon 600
- 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 Solid Cancer kit prepares sequencing libraries for genotyping KRAS, NRAS and BRAF 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
KRAS and NRAS, in the hotspot codons 12, 13, 59, 61, 117 and 146, and BRAF at codon 600. The manufacturer catalogue number is RDNGS0001.
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 RDNGS0001. 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.
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Application Protocol
Frequently paired with this kit
Other assays and instruments lab teams add alongside the Solid Cancer Kit (KRAS, NRAS, BRAF).
Add this kit to your quote
Tell us the platforms and throughput you’re planning for and our specialists will put together a clear proposal with the right documentation, no public pricing, just a considered answer.
