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黑麦B染色体显微切割和微克隆
MICRODISSECTION AND MICROCLONING OF THE B CHROMOSOME OF RYE ( Secale cereale )
【摘要】 显微分离出两条黑麦B染色体,经蛋白酶K处理,Sau3AI酶切后,用LA-PCR(LinkerAdapterPCR)方法进行扩增,得到0.2~2kb的DNA片段.Southern杂交分析结果表明,此扩增产物来源于黑麦B染色体.将部分PCR产物克隆到pUC19载体中,获得了5000个克隆.为构建B染色体DNA文库奠定了基础
【Abstract】 Two B chromosomes isolated from a metaphase cell of rye by microdissection, were digested with proteinase K and Sau3AⅠ, then the B chromosome DNA was amplified by LA PCR(Linker Adapter PCR).The PCR products are a smear ranged from 0.2 kb to 2 kb. The Southern hybridization showed that the PCR products were derived from the B chromosome of rye. Part of these PCR products were cloned into pUC19 and we obtained 5000 clones. This microdissection and microcloning method is a simple and general approach for constructing B chromosome DNA library.
【基金】 国家自然科学基金
- 【文献出处】 南开大学学报(自然科学版) ,JOURNAL OF NANKAI UNIVERSITY , 编辑部邮箱 ,1999年02期
- 【分类号】Q3-3
- 【被引频次】9
- 【下载频次】120