. The development of a cosmid map of chromosome 12p13. Sweet Briar College; Chromosomes. llYBRiniZATION. I lybridization using the digoxigenin system was found to produce successful results during this project. The three STS markers D12S950, D12S1987. and D12S1914 were labeled with PCR and were bound to DNA dot blots containing all the available cosmids. For each STS marker tested there were several very darkly labeled positive cosmids observed following incubation with x-ray film. Hybridization using psoralen biotin derivatives was not successful other than those procedures run as part
. The development of a cosmid map of chromosome 12p13. Sweet Briar College; Chromosomes. llYBRiniZATION. I lybridization using the digoxigenin system was found to produce successful results during this project. The three STS markers D12S950, D12S1987. and D12S1914 were labeled with PCR and were bound to DNA dot blots containing all the available cosmids. For each STS marker tested there were several very darkly labeled positive cosmids observed following incubation with x-ray film. Hybridization using psoralen biotin derivatives was not successful other than those procedures run as part of the control procedure. Control Hvbridizalion. A control hybridization was performed using DIG labeled template DNA. pBR328. supplied with the kit and self-labeled during PCR. Unlabeled pBR328 DNA was bound to a nylon membrane to form a dot blot. Probe concentration used for the procedure was 5 |iI7mL hybridization solution of which there was mL per 100 cm^ dot blot membrane. The control hybridization procedure revealed binding down to a 5x10 |ig/|aL concentration of template DNA (FIGURE 20). Mock hybridizations were carried out to determine the optimal probe concentration for experimental hybridizations. These hybridizations used small unlabeled pieces of membrane exposed to 1 |.iL/mL. 2 |jL/mL, 3 |.iL/mL. or 5 ^L/mL probe concentration during the overnight incubation. Following luminescence detection only the 5 concentration showed too high a background for use in experimental hybridizations. The 2 [iL/mL probe concentration was used for the experimental Fi(HiRi:2(). c iMiiidl Digoxigenin Hybridization using unlabeled pBR328 as the target DNA. DIG labeled pBR328 DNA was used as the probe. The probe produced a visible label for unlabeled DNA dilutions of lxlO'|.ig/|.iU 5x 10'Yig/|iL, \\\Q''"\<igl\.\L, and 5xlO"'ng''nL. 36. Please note that these images are extracted from scanned page images that may have been digitally enha
Size: 3074px × 813px
Photo credit: © Central Historic Books / Alamy / Afripics
License: Licensed
Model Released: No
Keywords: ., bookcentury1900, booksubjectchromo, booksubjectsweetbriarcollege
