Construction of fifteen human chromosome-specific DNA libraries from flow-purified chromosomes.
Construction of fifteen human chromosome-specific DNA libraries from flow-purified chromosomes.
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从流式纯化的染色体构建 15 个人类染色体特异性 DNA 文库。
DOI：
10.1159/000132301
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
VanDilla,MA
中科院分区：
文献类型：
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作者：
Fuscoe,JC;Clark,LM;VanDilla,MA
我们报告了15个人类染色体特异性DNA文库的构建。用流式分选法纯化中期染色体，提取DNA，用hindlll进行剪切，然后克隆到λ载体Charon 21a中。采用灵敏的微印迹杂交技术对克隆过程中的物理和生化步骤进行监测。利用这种方法，我们开发了一种高效的方案，用于从0.2-1.0 × 106排序的染色体中产生大量重组噬菌体。以下染色体的DNA被克隆：#4、6、8、9、11、13、14+15、16、17、18、19、20、21、22和Y.这些文库可供科学研究界使用，将在人类基因组的遗传分析中具有价值。
We report the construction of 15 human chromosome-specific DNA libraries. Metaphase chromosomes were purified by flow sorting and the DNA was extracted and cleaved withHindlllbefore cloning into the λ vector Charon 21 A. A sensitive miniblot hybridization method was used to monitor the physical and biochemical steps in the cloning procedure. Using this method, we have developed a highly efficient protocol for producing large numbers of recombinant phage from 0.2–1.0 × 106sorted chromosomes. DNA from the following chromosomes was cloned: #4, 6, 8, 9, 11, 13, 14+15, 16, 17, 18, 19, 20, 21, 22 and Y. These libraries are available to the scientific research community and will be valuable in the genetic analysis of the human genome.