MSI Analysis System, Version 1.2, Technical Manual
Literature # TM255
The MSI Analysis System, Version 1.2, is a fluorescent PCR-based assay for detection of microsatellite instability. Typically, MSI analysis involves comparing allelic profiles of microsatellite markers generated by amplification of DNA from matching normal and test samples, which may be MMR deficient. Alleles that are present in the test sample but not found in the corresponding normal samples indicate MSI. The MSI Analysis System includes fluorescently labeled primers for co-amplification of seven markers including five mononucleotide repeat markers (BAT-25, BAT-26, NR-21, NR-24 and MONO-27) and two pentanucleotide repeat markers(Penta C and Penta D). The mononucleotide markers are used for MSI determination, and the pentanucleotide markers are used to detect potential sample mixups and/or contamination. Internal lane size standards are added to the PCR samples to assure accurate sizing of alleles and to adjust for run-to-run variation. The PCR products are separated by capillary electrophoresis using an ABI PRISM® 310, 3100 or 3100-Avant Genetic Analyzer or Applied Biosystems 3130 or 3130xl Genetic Analyzer. The output data maybe analyzed with GeneMapper® Analysis Software to determine MSI status.
Summary of Changes: 8/14 Revision
The following changes were made to the 8/14 version of this document:
The module selected in Section 5.C, Step 4, was corrected to GS STR POP4 (1ml) F Module. Section 8.G was updated to reflect the fact that the panels and bins text files download package no longer includes an informational document and an example plot settings file.
Printed in USA. Revised 8/14.