High-throughput avian gender identification of Order Sphenisciformes

• Main Authors: Hui-Tzu Chiu, 邱慧慈
• Other Authors: Hsueh-Wei Chang
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/59465208968969667240

碩士 === 高雄醫學大學 === 生物醫學暨環境生物學研究所 === 100 === 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 and it is less high-throughtput. The purpose of my study is to design the primers or probes for the high-throughput gender identification of several species for the Order Sphenisciformes (penguins). I will introduce a PCR-melting curve analysis (PCR/MCA) to identify the gender of birds by genomic DNA, which is gel-free, quick, and inexpensive. Currently, I focus on the high-throughput gender identification for gentoo penguin (Pygoscelis papua) using the feathers samples. Using Griffiths'' P2/P8 primers, molecular sexing failed both in PCR/electrophoresis and in PCR/MCA of gentoo penguin. After the cloning and sequencing, the sex-specific primers for the CHD-Z and CHD-W genes of gentoo penguin, respectively, were redesigned for PCR/MCA. Using this specific primer set, 10 samples of gentoo penguin were examined simultaneously and the Tm peaks of CHD-Z and CHD-W PCR products were able to be distinguished. Therefore, the high-throughput avian molecular sexing technique was successfully applied to gentoo penguin.