PCR analyses of individual colonies.

DNA was processed from individually picked colonies (CFU-GEMM, CFC, and pre-B CFU) and analyzed by PCR for (1) quality of DNA preparation using the am (an autosomal gene) primers, (2) donor rather than recipient origin using Y chromosome-specific primers, and (3) transgene donor origin using both tEpoR and Neo primers. Shown is a representative sample of the PCR products from a male tEpoR tg donor (lanes 2, 5, 8, 11), male wild-type donor (lanes 3, 6, 9, 12), and female wild-type recipient (lanes 4, 7, 10, 13). Lane 1 contains Gibco 100-bp marker fragments. The primers used are indicated above the lane numbers. The bands corresponding to the primers are marked by arrows. Note, for example, the presence of the transgene-specific and Neo-specific bands in lanes 2 and 5, indicative of male-derived transgene-bearing colonies, and the absence of the Y-specific band in lane 13, indicative of a female-derived colony.