【摘要】 对猪(Susscrofa)基因组AFLP指纹图谱构建的各项条件进行了优化,包括双酶切消化、接头连接、预扩增和选择性扩增、凝胶电泳银染技术及荧光标记检测法等,建立了稳定的猪基因组DNA的AFLP指纹图谱构建方法,采用E32/T32引物组合在45个猪种基因组混合DNA中检测到28个多态标记。更多还原

【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.更多还原