. The development of a cosmid map of chromosome 12p13. Sweet Briar College; Chromosomes. Restriction Endonuclease Digestion and Agarose (lei Electrophoresis Southern Blot P. Figure 30. Roadmap of steps required to produce a physical map, or cosmid contig of a chromosome based on STS markers. Cosmids were generated through several cosmid preparations for use in making the dot blot and for restriction endonuclease digestions. The cosmids were also initially used in PCR testing for STS markers but after DIG was implemented as the more effective method of testing for marker sequences the template
. The development of a cosmid map of chromosome 12p13. Sweet Briar College; Chromosomes. Restriction Endonuclease Digestion and Agarose (lei Electrophoresis Southern Blot P. Figure 30. Roadmap of steps required to produce a physical map, or cosmid contig of a chromosome based on STS markers. Cosmids were generated through several cosmid preparations for use in making the dot blot and for restriction endonuclease digestions. The cosmids were also initially used in PCR testing for STS markers but after DIG was implemented as the more effective method of testing for marker sequences the template DNA was changed to K562 human DNA. PCR amplified and DIG labeled STS marker sequences were confirmed using agarose gel electrophoresis and then the labeled STS was used as a probe for hybridization to the dot blots of cosmid DNA. Hybridization with DIG labeled D12S950, D12S1987, and D12S1914 STS markers produced eleven, seven and eight strongly labeling positive cosmids, respectively. Restriction endonuclease digestions and agarose gel electrophoresis produced a way of analyzing the positive cosmids for overlaps when migration distances of the fragments were compared with those distances found for the fragments of known size. Arranging the determined fragment sizes in such a way as to maximize overlap between the cosmids allowed the production of two possible cosmid contigs for the region of the genome spanning the STS marker. 45. Please note that these images are extracted from scanned page images that may have been digitally enhanced for readability - coloration and appearance of these illustrations may not perfectly resemble the original Belk, Stephanie Heather.
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