TIPS & TECHNIQUES HOW TO: Next Generation Sequencing Sample Preparation for Ion Torrent™ Dr. Molly Accola is a Senior Laboratory Development Specialist at the University of Wisconsin Hospitals and Clinics. She works with next-generation sequencing (NGS) to develop assays to study variants in solid tumors. She recently talked with us about her work. Promega: Which NGS system do you use? When evaluating systems, how did you decide on a platform? Dr. Accola: That’s simple—it’s cost. Essentially we looked at the Illumina MiSeq and we looked at the Ion Torrent PGM™. The PGM and the OneTouch™ systems together were less than $100,000, which fit our budget. The workflow is also kitbased (Figure 1). Life Tech has a panel of oligos that’s prepackaged to target 50 oncology-associated genes. Their protocol, although a little tedious, is very straight forward and easy to use for a lab just starting NGS, so that’s why we chose the PGM. Promega: Could you describe the general process used in your lab of preparing and conducting NGS? Sequencing Obtain sample and purify nucleic acid Quantitate nucleic acid Prepare library Quantitate library Prepare Sequence sequencing template Analyze Data Figure 1. Workflow of sample preparation for NGS on Ion Torrent™. enews | March 2016 13170MA Pagina 13
Pagina 15Scoor meer met een webshop in uw sportbladen. Velen gingen u voor en publiceerden onderwijscatalogussen online.
Promega eNews March 2016 Nordic Lees publicatie 11025Home