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- Title
Utilization of Single Nucleotide Polymorphism-based Allele-specific PCR to Identify Shiikuwasha (Citrus depressa Hayata) and Calamondin (Citrus madurensis Lour.) in Processed Juice.
- Authors
MUSOU-YAHADA, Ayumi; HONJOH, Ken-ichi; YAMAMOTO, Kenta; MIYAMOTO, Takahisa; OHTA, Hideaki
- Abstract
To develop a method for the identification of shiikuwasha (Citrus depressa Hayata) and calamondin (Citrus madurensis Lour.), trnL-trnF and trnT-trnL intergenic spacer regions of their chloroplast DNA were amplified using PCR and the nucleotide sequences were determined. In each region, a single nucleotide polymorphism (SNP) site specific to the respective citrus species (shiikuwasha and calamondin) was found. For species discrimination using PCR, two forward primers containing the allele-specific SNP site at the 3'- end and a mismatched nucleotide at the 3rd base from the 3'-end were designed. The allele-specific forward primers specific to shiikuwasha and calamondin were respectively designated CiDeLF-F and CiMaTL-F. To confirm the specificity of the designed primers, PCR was carried out with DNA prepared from citrus peel or hand-squeezed juice as the template. Results showed that shiikuwasha and calamondin fruits and juices were identifiable by PCR using the allele-specific primers. Furthermore, this allele-specific PCR method can be applied to industrially processed and concentrated juice by amplifying DNA in advance.
- Subjects
SINGLE nucleotide polymorphisms; POLYMERASE chain reaction; GENE amplification; PRIMERS (Coating); ALLELES
- Publication
Food Science & Technology Research, 2019, Vol 25, Issue 1, p19
- ISSN
1344-6606
- Publication type
Article
- DOI
10.3136/fstr.25.19