Study of optimal method of Heterodera glycines sample preparation for molecular diagnostics

Abstract

In this study, it is optimized by the methods of preparing samples for the release of genetic material (DNA) from soybean cyst-forming nematode Heterodera glycines , which is necessary for accurate identification. Various modifications of ready-made reagent kits from Russian and foreign manufacturers were tested: DNA-Extrane-2, FitoSorb, TsytoSorb, ORB-GMO-A by Sintol LLC (Russia), Proba-GS, Proba-NK by AgroDiagnostika LLC, and QIAamp DNA Mini Kit by Qiagen (Germany). By testing the PCR method In real time, the sensitivity of Heterodera glycines total DNA release was tested using a test system consisting of forward and reverse primers and a modified probe: SCNrtF (5-AAATTCCAGGCCGCTATCTC-3), SCNrtR (5CGTGGACTGAACTGAACTGGACAAAG-3), SCNrtP (5-FAM/-TGGGCTGGGTGCTTCTAGAACTTTT-/ BHQ-1/-3).

Keywords

• soybean cyst nematode
• identification
• reagent kits
• dna extraction
• real-time pcr (rt-pcr)