Figure 2

Molecular mapping of MHC linked complement genes based on analysis of cosmid clones. A molecular map of MHC associated complement genes and steroid 21 hydroxylase loci based on EcoRI restriction pattern analysis of clones from a male sheep cosmid library. DNA (≈10 mg) from each clone was digested with EcoRI and size fractionated on 0.7% agarose gel and stained with ethidium bromide. The gel was then blotted onto Hybond N+ nylon membranes by vacuum assisted capillary blotting under alkaline conditions (0.2 M NaOH, 0.4 M NaCl). Following depurination, the membranes were then sequentially hybridised with probes in 5 × SSPE containing 7% SDS, 1% BSA and 5 mM EDTA and subsequently washed at elevated stringency (0.1 × SSC, 0.1% SDS, 65°C). Fragment sizes were determined from a 1 kb ladder (Bethesda Research Laboratories).