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Overview
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ChapterDx MLSTnext Listeria genotyping NGS assay is a unique and highly multiplex PCR assay that amplifies and analyzes 69 most polymorphic loci spanning L. monocytogenes genome. The assay is a single-tube and single-step PCR reaction, where amplification and indexing take places simultaneously in the same reaction. This assay provides high-resolution genotype information for L. monocytogenes and detect co-presence of different L. monocytogenes species from one sample. The assay is able to detect other Listeria species: L. grayi, L. innocua, L. ivanovii, L. marthii, L. seeligeri, and L. welshimeri. The high-resolution result allows tracking-and-tracing source of contamination. The data analysis software is included.
Features
· Analyze sequences for 69 polymorphic loci across L. monocytogenes genome
· Provide high-resolution genotypes for L. monocytogenes
· Detects all Listeria species including: L. grayi, L. innocua, L. ivanovii, L. marthii, L. seeligeri, and L. welshimeri
· Excellent tool for tracking-and-tracing the source of contamination
· Single-tube and single-step PCR
· User-friendly workflow and easy to automate
· Cost-efficient, and suitable for low to high-throughput scales
· Data analysis software includedPlease contact us at for specific academic pricing.
Background
ChapterDx Listeria Genotyping NGS assay consists of the following steps:
· DNA extraction (not provided)
· Multiplex PCR
· Pooling and purification
· Next-generation sequencing
In the PCR reaction, both amplification and indexing/barcoding of each specimen occurs simultaneously. After PCR reaction, amplicons for different samples are pooled in one tube. The following procedures deal with only one tube regardless of how many samples. After the cleanup step, the library is sequenced on an Illumina sequencing platform. The entire workflow takes about 24 hours, of which about 3 hours is benchwork/incubation time and 17 hours of sequencing. It takes less than half hour to complete data analysis.
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Overview