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Verification and applicability of endogenous reference genes for quantifying GM rice by digital PCR.

Authors
  • Deng, Tingting1
  • Huang, Wensheng2
  • Ren, Junan3
  • Ma, Xiuli1
  • Ge, Yiqiang4
  • Chen, Ying5
  • 1 College of Food Science and Nutritional Engineering, China Agriculture University, Beijing, 100083, People's Republic of China; Agro-product Safety Research Center, Chinese Academy of Inspection and Quarantine, Beijing, 100123, People's Republic of China. , (China)
  • 2 Agro-product Safety Research Center, Chinese Academy of Inspection and Quarantine, Beijing, 100123, People's Republic of China. , (China)
  • 3 Beijing Research Center for Agricultural Standards and Testing, Beijing Academy of Agriculture and Forestry Science, Beijjing, 100097, People's Republic of China. , (China)
  • 4 College of Food Science and Nutritional Engineering, China Agriculture University, Beijing, 100083, People's Republic of China; China Rural Technology Development Center, Beijing, 100045, People's Republic of China. Electronic address: [email protected] , (China)
  • 5 Agro-product Safety Research Center, Chinese Academy of Inspection and Quarantine, Beijing, 100123, People's Republic of China. Electronic address: [email protected] , (China)
Type
Published Article
Journal
Analytical Biochemistry
Publisher
Elsevier
Publication Date
Dec 15, 2019
Volume
587
Pages
113442–113442
Identifiers
DOI: 10.1016/j.ab.2019.113442
PMID: 31539524
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

To standardize the rice-specific quantification methods, the criteria of six genes of rice (gos9, PLD, SPS, RBE4, ppi-PPF and oriazain) were compared and evaluated by ddPCR. The results revealed that SPS, RBE4 and ppi-PPF were single copy genes per haploid genome and species specificity and stable among different rice cultivars, by employing Lectin gene of soybean as internal reference gene. The established ddPCR systems were precise and reliable with an absolute LOQ of 10-20 copies/reaction. Furthermore, the robustness of these three assays was verified by performing an intra-laboratory repeatability validation and the results showed that the three endogenous genes of rice could be quantitated repeatedly and precisely above the LOQ. These ddPCR methods can reliably quantified the GM content even if the content was low to 0.1%, which were much more reliable than the results from real-time PCR using the same primers and probes. Copyright © 2019 Elsevier Inc. All rights reserved.

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