Transformation of mammalian cells with recombinant DNA directly from Seaplaque agarose.
Transfection of African green monkey kidney cells directly with recombinant DNA excised from, but still present in, Seaplaque agarose after electrophoresis, is described. Efficiencies of transfection increased by 30% when the gel was present compared with transfection in the absence of the agarose. Extraction of the DNA from the gel was not necessary, thereby obviating a purification step and the concomitant losses. To generate recombinant molecules bacterial plasmid sequences are not necessary, thereby reducing considerably the size of the recombinant molecule and removing extraneous and deleterious sequences, e.g., "poison sequences." Linear or circular DNA molecules could be transfected in the melted and diluted agarose with the same ease as in its absence. Hence linear partial ligation products can be excised from the gel after electrophoresis to generate recombinant DNA molecules directly in mammalian cells.
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005822:Genetic Vectors; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004279:DNA, Viral