女贞不同种质材料基因组DNA的提取及RAPD引物筛选

Study on the Extraction of Genomic DNA and Screening of RAPD Primer in Different Germplasm Materials of Ligustrum lucidum Ait.

[目的]提取女贞不同种质材料的基因组DNA,并筛选RAPD引物。[方法]采用改良CTAB法提取女贞基因组DNA,并以此种方法提取的女贞基因组DNA为模板,对RAPD引物进行PCR扩增,筛选有效引物。[结果]筛选出21个多态性丰富、条带清晰且重复性好的有效引物,经检测所获得的基因组DNA条带清晰,且OD260/OD280在1.8左右。用筛选出的21个有效引物对126份女贞种质材料进行RAPD-PCR扩增,均可获得带型丰富且清晰可辨的DNA指纹图谱。[结论]该方法为快速和准确地应用RAPD方法分析女贞种质材料的遗传多样性提供了依据。

[ Objective ] To extract genomic DNA in different germplasm materials of Ligustrum lucidum and select RAPD primers. [ Method ] Genomic DNA was extracted by improved CTAB method, and with genomic DNA as a template, PCR amplification was conducted on RAPD primers. [ Result ] 21 effective primers with abundant polymorphism, clear bands and good repeatability were obtained. After testing the privet genomic DNA bands were clear, and the OD26o/OD2so was about 1.8. 21 selected effective primers were used in the RAPD-PCR amplification for 126 germplasm materials of L. lucidum. From all of which the abundant and distinct DNA fingerprinting could be obtained. [ Conclusion] The study will provide a reference for the rapid and accurate utilization RAPD method in analyzing genetic diversity of the germplasm materials of L. lucidum.