NDLI: High-throughput avian molecular sexing by SYBR green-based real-time PCR combined with melting curve analysis

Content Provider	Springer Nature : BioMed Central
Author	Chang, Hsueh-Wei Cheng, Chun-An Gu, De-Leung Chang, Chia-Che Su, San-Hua Wen, Cheng-Hao Chou, Yii-Cheng Chou, Ta-Ching Yao, Cheng-Te Tsai, Chi-Li Cheng, Chien-Chung
Abstract	Background Combination of CHD (chromo-helicase-DNA binding protein)-specific polymerase chain reaction (PCR) with electrophoresis (PCR/electrophoresis) is the most common avian molecular sexing technique but it is lab-intensive and gel-required. Gender determination often fails when the difference in length between the PCR products of CHD-Z and CHD-W genes is too short to be resolved. Results Here, we are the first to introduce a PCR-melting curve analysis (PCR/MCA) to identify the gender of birds by genomic DNA, which is gel-free, quick, and inexpensive. Spilornis cheela hoya (S. c. hoya) and Pycnonotus sinensis (P. sinensis) were used to illustrate this novel molecular sexing technique. The difference in the length of CHD genes in S. c. hoya and P. sinensis is 13-, and 52-bp, respectively. Using Griffiths' P2/P8 primers, molecular sexing failed both in PCR/electrophoresis of S. c. hoya and in PCR/MCA of S. c. hoya and P. sinensis. In contrast, we redesigned sex-specific primers to yield 185- and 112-bp PCR products for the CHD-Z and CHD-W genes of S. c. hoya, respectively, using PCR/MCA. Using this specific primer set, at least 13 samples of S. c. hoya were examined simultaneously and the Tm peaks of CHD-Z and CHD-W PCR products were distinguished. Conclusion In this study, we introduced a high-throughput avian molecular sexing technique and successfully applied it to two species. This new method holds a great potential for use in high throughput sexing of other avian species, as well.
Related Links	https://bmcbiotechnol.biomedcentral.com/counter/pdf/10.1186/1472-6750-8-12.pdf
Ending Page	8
Page Count	8
Starting Page	1
File Format	HTM / HTML
ISSN	14726750
DOI	10.1186/1472-6750-8-12
Journal	BMC Biotechnology
Issue Number	1
Volume Number	8
Language	English
Publisher	BioMed Central
Publisher Date	2008-02-12
Access Restriction	Open
Subject Keyword	Applied Microbiology Biotechnology Biochemical Engineering Genetic Engineering Plant Breeding Biomedical Engineering Polymerase Chain Reaction Polymerase Chain Reaction Product Polymerase Chain Reaction Reaction Melting Curve Analysis Regular Polymerase Chain Reaction Plant Breeding/Biotechnology Biomedical Engineering/Biotechnology
Content Type	Text
Resource Type	Article
Subject	Biotechnology
Journal Impact Factor	3.5/2023
5-Year Journal Impact Factor	3.2/2023

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