Construction of human gene libraries from small amounts of peripheral blood: analysis of beta-like globin genes.
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.
D001483:Base Sequence; D002648:Child; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D005914:Globins / Q000235:genetics*; D006720:Homozygote; D006801:Humans; D008297:Male; D008970:Molecular Weight; D013789:Thalassemia / Q000235:genetics*
D005914:Globins; D004247:DNA