Development of Genomic DNA Certified Reference Materials for Genetically Modified Rice Kefeng 6

Area of Science:

• Biotechnology
• Genetics
• Analytical Chemistry

Background:

• Accurate genetically modified organism (GMO) detection relies on certified reference materials (CRMs) for result comparability and traceability.
• Existing CRMs may lack sufficient accuracy or specific application for certain GM crops like rice Kefeng 6.

Purpose of the Study:

• To develop and characterize genomic DNA (gDNA) certified reference materials (CRMs) for GM rice Kefeng 6.
• To establish accurate and traceable quantitative reference points for GMO detection in rice.

Main Methods:

• Development of gDNA CRMs from GM rice Kefeng 6 leaves.
• Characterization using KF6/PLD duplex digital PCR across eight qualified laboratories.
• Assessment of homogeneity, stability, and freeze-thaw cycles.

Main Results:

• Certified property values for Kefeng 6 gDNA CRMs: copy number ratio (1.03 ± 0.04) and copy number concentration ((1.60 ± 0.11) × 10^5 copies/μL).
• Demonstrated good between-vial homogeneity at 2 μL sample intake.
• Recommended transport below 25 °C (cold chain preferred), with a shelf life of at least 12 months and a limit of 10 freeze-thaw cycles.

Conclusions:

• The developed Kefeng 6 gDNA CRMs offer accurate property values with combined uncertainties, serving as user-friendly calibrators.
• These CRMs enhance the reliability and standardization of GM rice Kefeng 6 inspection and monitoring.
• Contributes to a robust copy number-based reference system for GMO detection.