ORIGINAL ARTICLE
 
HIGHLIGHTS
  • One nematode can be used for DNA extraction by freezing treatment before extraction.
  • The commercial kit method is recommended for quick results of nematode DNA extraction.
  • CTAB and SDS methods are recommended if the time is available and costs are limited.
KEYWORDS
TOPICS
ABSTRACT
The root-knot nematode Meloidogyne graminicola is an economically important pest in rice production. The identification of a nematode species is an important basis in nematode management to reduce yield losses by extracting nematode DNA as an early step in molecular identification. This study aimed to investigate the optimal extraction method and number of M. graminicola for nematode genomic analysis based on PCR (polymerase chain reaction) and Sanger sequencing. The DNA extraction methods used in this study were the CTAB, SDS, and commercial kit (GeneAidTM Tissue/Blood DNA Mini Kit). The results revealed that the three DNA extraction methods could be used to analyze the nematode genomics based on PCR and Sanger sequencing using one nematode, both in a second-stage juvenile and a female, equipped with the process of nematode destruction by freezing. This finding was shown by the amplification of all DNA templates with Mg-F3 and Mg-R2 primers through PCR with a size of 370 bp, while Sanger sequencing obtained 372 bp.
ACKNOWLEDGEMENTS
The authors would like to thank Gadjah Mada University for research funded by RTA (Rekognisi Tugas Akhir-Student Final Project Recognition) with Contract No. 3550/UN1.P.III/Dit-Lit/PT.01.05/2022.
RESPONSIBLE EDITOR
Renata Dobosz
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
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