摘要
[目的] 筛选豆象干标本基因组DNA的有效提取方法.[方法] 以口岸截获时间不等的豆象干标本为材料,在传统方法上加以改进,对从干标本中提取豆象基因组DNA的方法进行了研究.将其所提取的基因组DNA的纯度和浓度与CTAB法和SDS法进行了比较,并对DNA进行了完整性及PCR验证.[结果] 与CTAB法和SDS法相比,豆象干标本提取方法提取的DNA OD260/OD280值均在1.7~1.9,且浓度合适.DNA完整性及PCR验证结果表明,豆象干标本提取方法比较适合后续PCR试验的开展.[结论] 豆象干标本提取方法适合该研究中豆象干标本基因组DNA的提取,可推广到检验检疫工作中截获豆象的检疫鉴定中,以缩短检验检疫工作周期,提高检验检疫工作准确性.
Abstract
[Objective] To select the effective DNA extraction method for dried samples of bean weevil insects.[Method] The method for extracting genome DNA was studied on the basis of traditional methods by using dried bean weevil insects of different intercepted times as materials.The purity and concentration of the extracted genomic DNA samples were detected and compared with CTAB and SDS method, then the extracted genomic DNA samples were further verified by the DNA integrity and PCR identification.[Result] Compared with CTAB and SDS method, the extracted genomic DNA samples had appropriate OD260/OD280 values between 1.7-1.9 and appropriate concentrations.The DNA integrity and PCR identification results showed that the genomic DNA extraction method for dried bean weevil samples was more suitable for subsequent PCR experiments.[Conclusion] Dried bean weevil DNA extration method is suitable for extrating bean weevil insects'genomic DNA.This method can be extended to the quarantine and identification of intercepted bean weevil samples in CIQ work, in order to shorten the period and improve the accuracy of CIQ work.
基金项目
山东出入境检验检疫局科技项目(SK201628)
山东出入境检验检疫局科技项目(SK201305)
烟台市科技发展计划项目(2008324)
国家质量监督检验检疫总局科技项目(2016IK313)
国家质量监督检验检疫总局科技项目(2015IK200)
国家科技期刊平台
NSTL
维普
万方数据