Vor-Ort-molekulare Erkennung von bodenbürtige Phytopathogene mit einem tragbaren Echtzeit-PCR-System

Hi my name's Joseph DeShields, I'm with the Department of Plant Pathology at Washington State University and today I'll be showing you an onsite detection protocol for detecting soil-borne diseases in soil.Onsite diagnosis of plant diseases in the field is pivotal for effective disease management and can ultimately reduce secondary spread of pathogens in the field.Soil-borne pathogens are particularly difficult to diagnose due to the large and complex soil environments and this can make understanding soil-borne diseases difficult.Polymerase chain reaction, otherwise known as PCR, is currently the most widely-accepted sensitive and accurate approach to detecting plant pathogens in the field.The following protocol demonstrates a sensitive and onsite method for detecting soil-borne pathogens comprised of a magnetic bead-based DNA extraction, PCR, which can be done using a portable PCR instrument, and data analysis, which can be done remotely, using a laptop computer.Magnetic bead-based DNA extraction.The DNA and RNA extraction kit was prepared and used per the manufacturer's instructions with slight modifications.All reagents were stored at room temperature and the prepared Proteinase K was stored at a 20