Poncz M, Solowiejczyk D, Harpel B, Mory Y, Schwartz E, Surrey S
Hemoglobin. 1982;6(1):27-36. doi: 10.3109/03630268208996930.
We describe a rapid procedure for constructing cloned human genomic libraries from small amounts of peripheral blood. High molecular weight DNA is isolated from 5-20 ml peripheral blood, partially cleaved with Eco R1, and 8-22 kb fragments are cloned using bacteriophage Charon 4A and suitable E. coli host. Using the approach we have isolated and characterized several non-alpha globin clones from a Kurdish Jew with homozygous beta thalassemia. The ability to isolate suitable amounts of high molecular weight DNA from peripheral blood provides a relatively simple means of constructing human gene libraries representing a variety of hemoglobin disorders.
我们描述了一种从少量外周血构建克隆人基因组文库的快速方法。从5 - 20毫升外周血中分离高分子量DNA，用Eco R1进行部分切割，使用噬菌体Charon 4A和合适的大肠杆菌宿主克隆8 - 22 kb的片段。通过这种方法，我们从一名患有纯合β地中海贫血的库尔德犹太人中分离并鉴定了几个非α珠蛋白克隆。从外周血中分离出适量高分子量DNA的能力为构建代表各种血红蛋白疾病的人类基因文库提供了一种相对简单的方法。
