（1.江西农业大学江西省动物生物技术重点实验室，南昌 330045； 2.Department of Pathology, University of Cambridge, United Kingdom）
AFLP Markers for Pig Genomic DNA Fingerprinting
Ren Jun1 Huang Lusheng1** Gary Evens2 Gao Jun1 Ai Huashui1 Chen Kefei1 Ding Nengshui1 Deng Suhua1
(1. Jiangxi Provincial Key Laboratory for Animal Biotechnology, Jiangxi Agricultural University, Nanchang 330045, China； 2. Department of Pathology, University of Cambridge, United Kingdom)
Abstract：Abstract: The optimization of AFLP technique for porcine（Sus scrofa）genomic DNA fingerprinting was studied, including enzyme digestion, adapter ligation, preamplification, selective amplification, denatured polyacrylamide gel electrophoresis, silver staining and multicolor fluorescent detection. The Eco RⅠ+AAC/TaqⅠ+AAC selective primer combinations labeled by IRD 800 and IRD 700 fluorescent dyes were used. Gel running was performed in both Licor 4200 DNA sequencer and 4.5% PAGE(2600 V, 45 W, 50 ℃), and the electrophoresis images were analysed by the AFLP QuantarTM Pro software of Keygene(the Netherland). The result showed that the template DNA concentrations did not affect the AFLP quality, templates with 250 ng, 500 ng and 1 000 ng DNA had the same AFLP fingerprinting. And 28 polymorphic markers were detected in the same pooled genomic DNA of 45 pig breeds (populations) by E32/T32 primer combinations.
