Eligene Colorectum

Kit: EliGene® Colorectum NGS Kit (ref.: 90066-NGS)
Package size:
 24 reactions
Accessories: EliGene® Adaptor-IL NGS Kit (ref.: 90069-NGS, 96 reactions)
EliZyme Library Quant. Kit (ref.: EZ4815 - 500 - 1000 rxns)
MiSeq Reagent Nano Kit (Ref.: MS-103-1001, 15 reactions) or MiSeq Reagent Micro Kit (Ref.: MS-103-1002, 50 reactions)
Extraction kit EliGene FFPE Tissue DNA kit (ref.: 415050 - 50 reactions)
Specimens: blood, FFPE material
Compatible instruments:
 MiSeq (Illumina)
CE certification: yes
Detection technology: multiplex PCR, Next Generation Sequencing
Description
EliGene® Colorectum NGS kit is intended for multiplex PCR of target regions of KRAS (exons 2,3 and 4), NRAS (exons 2,3 and 4) and BRAF genes (exon 15) allowing detection of polymorphisms included in the amplified regions.
Principle of the method
EliGene® NGS kits enable multiplex PCR amplification of all required target regions of the gene(s) of interest. The recommended amount of DNA for each multiplex PCR reaction is between 20 and 100 ng of DNA derived from formalin-fixed paraffin-embedded (FFPE) material or from blood. Next, the resulting amplicons are barcoded with MID sequences, and mixed per sample. This sample is quantified and sequenced using a NGS instrument Illumina for massively parallel sequencing (MPS) according to the manufacturer’s instructions. The resulting sequence reads are subsequently analyzed by comparison with the reference sequence of the targeted gene(s) to identify variant positions.
References:
De Leeneer K, De Schrijver J, Clement L, Baetens M, Lefever S, et al. (2011) Practical Tools to Implement Massive Parallel Pyrosequencing of PCR Products in Next Generation Molecular Diagnostics. PLoS ONE 6(9).