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United States Department of Agriculture

Agricultural Research Service

Title: Directed Cosmid Isolation for Physical Assignment by Fish: Pcr-Based Iterative Screening of Random Genomic Libraries

item Heaton, Michael
item Lopez, Nestor
item Smith, Timothy
item Kappes, Steven
item Beattie, Craig

Submitted to: Animal Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 17, 1997
Publication Date: N/A

Interpretive Summary: Genetic linkage maps have recently been constructed for a variety of animal species used in agriculture. The efficient use of these maps for improving animal breeds depends on their quality. A new method was developed for determining the overall quality of genetic linkage maps and for identifying regions of the animal genome that are not covered by current maps. The method was demonstrated by using bovine chromosome 28 as an example to estimate genome coverage. Since this method is broadly applicable, its use allows information from the gene-rich human map to be efficiently applied to lower resolution livestock maps.

Technical Abstract: A directed PCR-based iterative screening protocol was developed to isolate cosmids containing microsatellite (ms) markers linked to chromosomal regions of interest, such as those near the ends of linkage groups or quantitative trait loci. This method was optimized for large-scale screening of total genomic libraries and used to purify bovine cosmids that anchor the ends of the bovine linkage group 28 (BTA28). Cosmids containing ms markers RBP3 and BMS2060 were purified for fluorescence in situ hybridization and assigned to 28q18-q19 and 28q12, respectively. These assignments indicated that approximately 73% of BTA28 (90% excluding the centromere) is covered by the current linkage map. Since this method is applicable to any target gene sequence suitable for PCR amplification, it may be extended to comparative mapping of genes.

Last Modified: 4/22/2015