Figure 3.

Pipeline fidelity and efficiency. (a,b) Fidelity of 96-well recombineering. Typical results of a 96-well-format recombineering experiment are shown (a). In the control experiment (b), the plate was inverted so that the BAC and the targeting cassettes did not match, resulting in virtually no background growth and thereby indicating that growth in selective medium is only a result of the intended recombination event and does not occur from random cassette integration. (c) Western blot analysis of 67 transgenic cell pools using an antibody to GFP. Lane 1 represents a wild-type HeLa cell control. For 57 cell pools (marked in green), a band of the expected size was detected (marked with *). For three genes, the band pattern did not represent the expected size (marked in yellow: lanes 3, 11, 57). For seven of the pools, no western blot band was observed (marked in red). In lane 9 (mUBE1), three bands matching the size of the human splice forms were detected. Numbers at left indicate molecular masses in kDa. More information on the tagged genes can be found in Supplementary Table 1 online.