Optimized MOL-PCR for Characterization of Microbial Pathogens.

Optimized MOL-PCR for Characterization of Microbial Pathogens. Curr Protoc Cytom. 2016;75:13.15.1-13.15.15 Authors: Wuyts V, Roosens NH, Bertrand S, Marchal K, De Keersmaecker SC Abstract Characterization of microbial pathogens is necessary for surveillance, outbreak detection, and tracing of outbreak sources. This unit describes a multiplex oligonucleotide ligation-PCR (MOL-PCR) optimized for characterization of microbial pathogens. With MOL-PCR, different types of markers, like unique sequences, single-nucleotide polymorphisms (SNPs) and indels, can be simultaneously analyzed in one assay. This assay consists of a multiplex ligation for detection of the markers, a singleplex PCR for signal amplification, and hybridization to MagPlex-TAG beads for readout on a Luminex platform after fluorescent staining. The current protocol describes the MOL-PCR, as well as methods for DNA isolation, probe design, and data interpretation and it is based on an optimized MOL-PCR assay for subtyping of Salmonella Typhimurium. © 2016 by John Wiley & Sons, Inc. PMID: 26742655 [PubMed - in process]
Source: Current Protocols in Cytometry - Category: Molecular Biology Tags: Curr Protoc Cytom Source Type: research