Matrix-based DNA Extraction for Targeted Next-Generation Sequencing on Decontaminated Sputum SamplesJennifer Williams *1, Janré Steyn *1, Emilyn Costa Conceição *1, Felicia Bernita Wells 1, Melanie Grobbelaar 1, Nabila Ismail 1, Yonas Ghebrekristos 1,2, Christoffel Johannes Opperman 1,2,3,4, Sarishna Singh 1,2, Jason Limberis 5, Fahd Naufal 5, Brendon Coenrad Mann 1, Rebecca E. Colman 6,7, Timothy Rodwell *6,7, Robin Mark Warren *1, John Z. Metcalfe *5
1Division of Molecular Biology and Human Genetics, SAMRC Centre for TB Research, DSI-NRF Centre of Excellence for Biomedical TB Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, 2National Health Laboratory Service, Green Point Tuberculosis Laboratory, 3Division of Medical Microbiology, Department of Pathology, University of Cape Town, 4Division of Medical Microbiology, Department of Pathology, University of Cape Town, 5Division of Pulmonary and Critical Care Medicine, Zuckerberg San Francisco General Hospital and Trauma Centre, University of California, San Francisco, 6Department of Medicine, University of California, San Diego, School of Medicine, 7Foundation for Innovative New Diagnostics (FIND), Campus Biotech
We present an optimized method for DNA extraction from decontaminated sputum using a matrix-based extraction method combined with a magnetic bead purification for downstream targeted next-generation sequencing of Mycobacterium tuberculosis.
