APPLICATION OF 3 DIFFERENT METHODS TO ANALYZE FIBER-FISH RESULTS OBTAINED USING 4 LAMBDA-CLONES FROM THE PORCINE MHC-III REGION

Four lambda clones (lambda G11, lambda C4, lambda G14 and lambda G17) from the porcine MHC class ill region were labelled with either biotin -14-dATP or digoxigenin-11-dUTP and hybridized in two different combin ations to DNA fibres. The latter were prepared from agarose-embedded p orcine peripheral lymphocytes lysed with proteinase K, then spread and fixed on poly-L-lysine-coated slides. Hybridization signals thus obta ined confirm the order of the clones previously reported using pulsed- field gel electrophoresis (PFGE). Measurements of probe sizes, gap dis tances between probes and total length of DNA encompassing the probes were made. Three different methods, namely relative length, Watson-Cri ck standard and probe size standard-based conversions, were used to es timate the parameters in kilobases. These methods of data conversion w ere compared with each other and with the available PFGE data, and the ir utility and accuracy were evaluated.