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The protein truncation test was used to search all regions of the patient's APC gene for germline mutations [R. van der Luijt et al., Genomics 20, 1 (1994)]. This assay provided evidence of a mutation in region G of APC exon 15. This region was specifically amplified from genomic DNA by polymerase chain reaction (PCR), and the PCR product was sequenced in forward and reverse orientation.
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The reference for the cY97 probe is J. Wolfe et al., J. Mol. Biol 182, 477 (1985). The pDP105 probe was obtained from the American Type Culture Collection (Rockville, MD) [M. Andersson, D. C. Page, A. de la Chapelle, Science 233, 786 (1986)].
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The reference for the cY97 probe is J. Wolfe et al., J. Mol. Biol 182, 477 (1985). The pDP105 probe was obtained from the American Type Culture Collection (Rockville, MD) [M. Andersson, D. C. Page, A. de la Chapelle, Science 233, 786 (1986)].
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DMA probes were labeled with biotin-14-adenosine triphosphate (Boehringer Mannheim, Lewes, UK) with the use of a Bionick kit (Life Technologies, Paisley, UK). The DNA was purified by passage through a Sephadex G50 column and ethanol precipitation NISH was performed as previously described (H. M J. Kerstens, P. J. Poddighe, G. J. M. Hanselaar, Histochem. Cytochem. 42, 1971 (1994)]. Signal was detected immunohistochemically with (i) monoclonal mouse antibody to biotin (1:20) (Dakopatts, High Wycombe, UK), (ii) biotinylated rabbit immunoglobulins to mouse immunoglobulins (1:200) (Dakopatts), and (iii) streptavidin-HRP (1:400) (Dakopatts).
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Dysplasia is defined as "cells having undergone proliferation and atypical cytological alterations involving cell size, shape, and organization" [R. S. Cotran, V. Kumar, S. L. Robbins, Robbins Pathological Basis of Disease (W. B. Saunders, Philadelphia, PA, 1989)].
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-9. Thus, the model suggests that the collision hypothesis cannot account for the observed data.
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Pyke, R.1
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15844422912
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note
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For mixed (XO/XY) adenomas, mean width = 5.93 crypts (n = 13); and for all adenomas, mean width = 6.82 (n = 285) and SEM = 0.554. The width of mixed adenomas does not differ significantly from that of the general polyp population (normal distribution, two-tailed test, P > 0.1).
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34
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note
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We thank P. Sasieni for assistance with the statistical analysis and A. Rowan for performing the protein truncation test.
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