Isolation of native DNA fragments containing structural genes at the beginning, in the middle or at the end of the coding strand.
A method for the isolation of the DNA fragments containing structural genes at the beginning, in the middle and at the end of the coding strand is described. It involves mechanical or enzymatic shearing, exonuclease treatment, hybridization with mRNA and subsequent retention of DNA-mRNA complex on poly(U)-Sepharose. Hybridization of the DNA fragments with mRNA takes place with high specificity and isolated material is enriched in structural genes. Applications of this method for gene isolation are discussed.
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis; D002846:Chromatography, Affinity; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D005092:Exonucleases; D005796:Genes; D008099:Liver / Q000032:analysis; D008722:Methods; D051379:Mice; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000302:isolation & purification
D012333:RNA, Messenger; D004247:DNA; D005092:Exonucleases; D004262:DNA Restriction Enzymes