Molecular techniques are crucial for research on nematology, and the
processing of samples is essential for their long-term storage prior to analysis. This
chapter focuses on DNA extraction, various DNA markers, and polymerase change
reaction (PCR). To ensure the preservation of nematodes for extended periods, DESS
can be utilized. Moreover, the quality of extracted DNA is vital for proper PCR
processing. This chapter offers an overview of nematode isolation and the different
techniques for DNA extraction. The utilization of molecular markers presents
numerous benefits when studying the genetic makeup of nematode populations. Among
these markers, sequence characterized amplified region (SCAR) stands out for its
exceptional value in identifying nematodes belonging to the Meloidogyne species.
Amplified fragment length polymorphism (AFLP), on the other hand, is a technique
that can be employed to analyze the genetic variability of nematodes. It is important to
note that these markers are primarily used for plant-parasitic nematodes, which have a
significant impact on the economy. This chapter focuses on the most commonly
utilized marker for nematodes. After extracting DNA, the next step involves amplifying
the target genes through PCR. Various primers, including rDNA and mtDNA, are
available for nematode genes and serve useful taxonomic purposes. However, precise
primer design is critical for achieving accurate nematode diagnosis. Both DNA and
primers must be of high quality to ensure successful PCR. Eco-friendly standard dyes
like SafeView can be employed to visualize PCR products. This chapter offers a
comprehensive guide to PCR processing and DNA visualization techniques.
Keywords: Chelex, DNA extraction, DNA marker, Genetic diversity, Primer, Sequencing, Thermocycler, Visualization.
