Identification of PDGF-D transgenic mice by Southern Blot

Objective To identify the PDGF D transgenic c57 mice by Southern Blot.Methods:Transgenic mice were generated by microinjection of purified transgenic Alpha MyHC PDGF D vector into the pronuclei of fertilized eggs.Transgenic mouse genomic DNA was isolated from mice tails.After identification with PCR,the positive mice were further identified with Southern Blot.10 μg of the genomic DNA was digested with EcoRI,followed by fractionation through a 0 8% agarose gel and transferred onto a nylon membrane.The hGH PolyA probe was produced by PCR.After the hybridization with a 32 P labeled random primed hGH PolyA probe,the membranes were subjected to autoradiography for about one month.Results:The results of Southern Blot indicated that there is only one positive PDGF D transgenic mouse in these 20 mice.