Construction of Human Gene Libraries from Small Amounts cf Peripheral Blood: Analysis of β-Like Globin Genes

M. Poncz Division of Hematology The Children's Hospital of Philadelphia Departments of Pediatrics and Human Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104

D. Solowiejczyk Division of Hematology The Children's Hospital of Philadelphia Departments of Pediatrics and Human Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104

B. Harpel Division of Hematology The Children's Hospital of Philadelphia Departments of Pediatrics and Human Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104

Y. Mory The Weizmann Institute of Science, Rehovot, Israel

E. Schwartz Division of Hematology The Children's Hospital of Philadelphia Departments of Pediatrics and Human Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104

S. Surrey Division of Hematology The Children's Hospital of Philadelphia Departments of Pediatrics and Human Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104

Abstract

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 a suitable E.coli host. Using this approach we have isolated and characterized several non-a globin clones from a Kurdish Jew with homozygous β 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.

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Construction of Human Gene Libraries from Small Amounts cf Peripheral Blood: Analysis of β-Like Globin Genes

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Construction of Human Gene Libraries from Small Amounts cf Peripheral Blood: Analysis of β-Like Globin Genes

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