OncoMate™ MSI Dx Analysis System Technical Manual

Instructions for Use of Product(s)

Literature # TM543

The OncoMate™ MSI Dx Analysis System encompasses a complete workflow for MSI determination, from DNA extraction to data analysis. First, DNA is extracted from FFPE colorectal tissue samples (normal and tumor from the same patient) using the Maxwell® CSC DNA FFPE Kit and Maxwell® CSC Instrument. Double-stranded DNA (dsDNA) is then quantified using a fluorescence-based dsDNA quantification system of your choice. Next, amplification products are generated through multiplex PCR amplification of DNA microsatellite markers using the OncoMate™ MSI Dx Analysis System amplification kit (Cat.# MD2140). The PCR products are then mixed with Hi-Di™ Formamide and Size Standard 500 and heat-denatured. The resulting single-stranded DNA fragments are separated by size and detected via fluorescence using an Applied Biosystems® 3500 Dx Genetic Analyzer. Following capillary electrophoresis (CE), allele sizes from the CRC tumor DNA and the normal DNA are calculated and compared for each of the microsatellite markers using OncoMate™ MSI Dx Interpretive Software. If the length of two or more of the five mononucleotiderepeat marker alleles is changed by ≥2.75 base pairs (bp), the tumor is classified as MSI-H; if the allele length is changed for only one marker, or if the difference in allele lengths at the five markers is <2.75bp, the tumor is classified as microsatellite stable (MSS). The sizes of the Penta C and Penta D pentanucleotide-repeat marker alleles are compared as an identity check between the normal and tumor DNA samples.

This product is only available in the United States.