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Molecular biology-based lab methods using plant nematodes
Kris Lambert: University of Illinois
<div>The identification of plant parasitic nematodes in a laboratory class setting can be challenging. Often high-resolution microscopic equipment required for careful nematode observations are not available. While counter intuitive, the use of molecular biological techniques in nematode identification is often simpler, more accurate and offers students the opportunity to combine classical nematology techniques with more modern laboratory methods. By mixing a combination of “hands-on” nematode wrangling, micro-scale DNA extraction and computational analysis of nematode DNA sequences, the often tedious task of plant nematode identification can become a highly interactive experience for undergraduate plant pathology students. To successfully use molecular methods in identifying plant nematodes, access to a thermocycler and basic agarose gel electrophoresis equipment, common in many research laboratories, is necessary. Having DNA sequence files for student practice is also useful. For molecular methods to function smoothly in a plant pathology laboratory, it is critical that all methods are as streamlined as possible. Simple single-tube DNA extractions and PCR reactions are conducted by the students and then, while the thermocyclers are running, a discussion on how the data is analyzed is conducted. Students are provided with a list of web sites that host free bioinformatics tools so they can analyze sequence data and conduct homework assignments. The use of locally extracted nematodes from student-collected samples can provide a useful survey of common plant nematodes in the area and raise awareness of nematode ecology, which can feedback in to the lecture portion of the class.</div>

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