Rapid two-stage polymerase chain reaction amplification of chromosomal DNA segments in lysates made from monolayer cultures attached to microcarrier beads.
We describe a method for the rapid two-stage amplification and detection by ethidium bromide staining of chromosomal nucleotide (nt) sequences in lysates made directly from anchorage-dependent cells attached to microcarrier beads. The procedure circumvents the need for cell detachment steps prior to analysis, facilitates the collection, transfer, and manipulation of the cells being studied, and makes unnecessary the use of Southern-blot hybridization for identification of specific nt sequences present in a small fraction of cells within a heterogeneous population.
Lin-Chao
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Department of Genetics, Stanford University School of Medicine, CA 94305-5120.
Brenner
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DG
Cohen
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D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D002460:Cell Line; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis*; D007763:Lac Operon; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004247:DNA