[PCR detecting homologous recombinants from ES cells].
PCR is an effective method for detecting homologous recombinants from ES cells, because there are different genomic DNA structure among non-recombinants, site specific recombinants and non-site specific recombinants. When we design suitable primers, we can analyze and identify site specific recombinants, non-site specific recombinants and non-recombinants. In this paper, we design two different pairs of primer (according to the genomic DNA patterns) for using PCR to detect the cells which resist to G418 and 6-TG after plasmid pRV4.0 transforming male mouse ES cells, we get the homologous recombinants quickly, simply and specifically. We test the reliability of PCR using southern blot hybridization.
Xue
Y
Y
Department of Biology, Peking University, Beijing.
Liu
X
X
Wa
H
H
D000818:Animals; D001483:Base Sequence; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013234:Stem Cells / Q000378:metabolism*