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On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
Summary February 23rd, 2018
Rapid and accurate detection of plant pathogens on-site, especially soil-borne pathogens, is crucial to prevent further inoculum production and proliferation of plant diseases in the field. The method developed here using a portable real-time PCR detection system enables on-site diagnosis under field conditions.
Transcript
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
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