摘要
以卷丹百合(Lilium lancifolium Thunb.)为试材,改良CTAB法提取DNA,通过对优化酶切、连接、预扩增、选择性扩增等试验条件筛选,初步建立适合卷丹百合的AFLP反应体系。结果表明,改良CTAB法提取的卷丹百合基因组DNA符合AFLP分析要求;最佳酶切时间为37℃酶切8 h;16℃连接过夜;预扩增产物稀释30倍为宜。利用该体系筛选出12对能获得稳定、清晰扩增图谱的引物组合,为卷丹百合遗传图谱构建及分子辅助育种奠定基础。
An AFLP reaction system was established with Lilium lancifolium Thunb. by optimizing some factors including enzyme digestion, ligation, pre-amplification and selective amplification. The results showed that the genomic DNA extracted by the improved CTAB method was fit for AFLP analysis. The digestion duration should be 8 h at 37 ℃. The ligation should be overnight at 16 ℃. The products of pre-amplification should be diluted by 30 times. The clear and stable amplification patterns could be obtained from 12 pairs of primer combinations with the optimized system. This paper might provide a method involving in gene mapping and marker-assisted breeding of Lilium lancifolium Thunb.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2013年第10期122-127,F0003,共7页
Journal of Northeast Agricultural University
基金
辽宁省教育厅基金项目(202053092)
关键词
卷丹百合
AFLP
反应体系
优化
Lilium lancifolium Thunb.
AFLP
reaction system
optimization
