摘要
研究构建了一个脐橙基因组DNA文库。以‘大山岛’脐橙幼叶为试材 ,CTAB法制备基因组DNA ,经CsCl密度梯度离心纯化 ,获得了大片段 ( >10 0kb)高纯度基因组DNA。以Sau3AI部分酶切 ,酶切片段 3’凹端不完全补平后与LambdaGEM 12XhoIHalf SiteArms连接 ,连接产物用Packagene Extract包装 ,所得噬菌体在KW2 51寄主上铺平板。文库经过扩增并保存。对该文库特性研究表明 ,该文库包含了 2 .15× 10 6个单个克隆 ,背景低于10 0pfu/ μgDNA ,插入片段平均大小约为 17kb ,证明是一个较为完整的脐橙基因组DNA文库。
A genomic DNA library of citrus( Citrus sinensis Osbeck)was constructed in the study.Citrus genomic DNA was prepared with the CTAB method,and purified by CsCl spin.After the genomic DNA was partially digested with Sau 3A I,the first two nucleotides of this site were filled with Klenow.Then the fragments were ligated with LambdaGEM 12 Xho I Half Site Arms.The ligation mixture was packaged using the Packagene extract and titrated on LB plates.A DNA library with 2.15×10 6 individual clones and a low background at 100pfu/ug DNA was obtained.The average length of inserts is about 17kb.The library was proved to be standard and representative.
出处
《园艺学报》
CAS
CSCD
北大核心
2000年第6期401-405,共5页
Acta Horticulturae Sinica
基金
国家自然科学基金资助项目!(39770 5 2 6
39730 340 )
