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| SUMMER SCHOOL - Module 2 |
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| Module 2/ Introduction to Oxford nanopore (MinION) and MiSeq (illumina) technologies: preparation of whole microbial genome, metagenome and amplicon libraries. Dates: June 30th- July 3rd 2020 Coordinator: Llúcia Martínez Priego (martinez_lucpri@gva.es) The Sequencing Service of Fundació per al Foment de la Investigació Sanitària i Biomèdica de la Generalitat Valenciana (FISABIO – Salut Pública) propose a basic course in Oxford nanopore (MinION) and MiSeq (illumina) technologies. The goal of this course is to teach the fundamentals of nanopore and illumina library preparation protocols and sequencing. The training will be focused on microbiology NGS applications: microbial whole genomic, metagenomics and 16S amplicon protocols. The course is addressed to professionals of microbiology services in hospitals, NGS laboratory technicians and other professionals in microbiology, health sciences and related fields who are interested in learning new techniques to approach the study of microbiota. The module will be practical and hands-on, and will teach participants to prepare two different libraries for Illumina platforms: 16S amplicon libraries for taxonomy analyses and whole genome amplification libraries for genomic or metagenomic analyses and one library protocol for Oxford nanopore MinION platform: 16S amplicon libraries. Brief theoretical sessions and practical hands-on sessions will be taught by FISABIO Sequencing Service staff. Official language: English or Spanish (depending of the student's origin) Module costs: 480 euros Instructors: Llúcia Martínez Priego PhD. Manager in the Sequencing Service of FISABIO-SP.
Tuesday June 30th: 09:15 – 10:00 An Introduction to NGS library preparation 10:00 – 10:30 Best practices in a NGS laboratory Break 11:00 – 12:00 DNA quantification and normalisation 12:00 – 13:00 Starting illumina 16S amplicon protocol: first amplicon PCR Lunch 14:30 – 16:30 Starting illumina microbial whole genomic protocol: DNA tagmentation and library clean up 16:30 – 17:30 Indexing both illumina protocols 09:00 – 10:00 Cleaning up both illumina protocols Llúcia and Griselda 10:00 – 10:30 Bioanalyzer set up for illumine libraries validation Llúcia and Griselda Break 11:00 – 11:30 Library quantification Llúcia and Griselda 11:30 – 13:00 Review of library validation results Llúcia Martínez Priego Lunch 14:30 – 16:30 Library normalization Llúcia Martínez Priego 16:30 – 17:30 Create Sample sheet for Illumina MiSeq RUN Llúcia Martínez Priego Thursday July 2nd: 09:15 – 10:30 An Introduction to Oxford nanopore technology Llúcia Martínez Priego Break 11:00 – 13:00 Nanopore 16S amplicon library preparation Llúcia and Griselda Lunch 14:30 – 16:30 Run ONT device and assess sequencing performance during the run Llúcia and Griselda Friday July 3rd: 09:00 – 10:45 Preparing the MiSeq and the loading cartridge Llúcia Martínez Priego Break 11:00 – 13:30 Loading libraries for sequencing Llúcia Martínez Priego 13:30 – 15:00 Course feedback and wrap up Llúcia Martínez Priego | ||
