Archive image from page 42 of The development of a cosmid. The development of a cosmid map of chromosome 12p13 . developmentofcos00belk Year: 1998 0P_O—P—0—P—0 I I I Figurf9. Digoxigenin-11-dUTP. The labeled STS marker probes will then bind with great specificity to cosmids that contain their complementary sequence. This hybridization is easily visualized by immunochemical detection of the hybridized probes. Anti-digoxigenin fragments conjugated to alkaline phosphatases bind CSPD, a chemiluminescence substrate, to produce luminescence on the nylon membrane of the dot blot.' The luminescenc
Archive image from page 42 of The development of a cosmid. The development of a cosmid map of chromosome 12p13 . developmentofcos00belk Year: 1998 0P_O—P—0—P—0 I I I Figurf9. Digoxigenin-11-dUTP. The labeled STS marker probes will then bind with great specificity to cosmids that contain their complementary sequence. This hybridization is easily visualized by immunochemical detection of the hybridized probes. Anti-digoxigenin fragments conjugated to alkaline phosphatases bind CSPD, a chemiluminescence substrate, to produce luminescence on the nylon membrane of the dot blot.' The luminescence can be recorded on film. It is possible to produce probes specific to a particular STS for use in hybridization to cosmid dot blots, by using primers specific for STS markers in the pi3 region of Chromosome 12 (FIGURE 10). ADVANTAGES TO USING DiGOXIGENINHYBRIDIZATION. The digoxigenin system has a much easier method of labeling, using PCR, compared to the long irradiation step and multiple extraction steps required in psoralen biotin labeling. Hybridization and chemiluminescent detection procedures are similar in both techniques. In the digoxigenin labeled control it is possible to detect a DNA concentration down to pg of homologous DNA using chemiluminescent detection. The first detectable results are visible after only fifteen to twenty minutes, depending on the strength of the signal, and are visible for up to two days, allowing for multiple exposures.' In contrast, chemiluminescent control labeling with psoralen biotin is less sensitive and will detect 50 pg of DNA within two or three hours of film exppsure. 20
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Keywords: 1990, 1998, archive, belk_stephanie_heather, book, bookauthor, bookdecade, booksubject, bookyear, chromosomes, drawing, historical, history, illustration, image, page, picture, print, reference, sweet_briar_college, vintage
