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Title: USE OF DIRECT IN SITU SINGLE COPY PCR TO DEVELOP COMPREHENSIVE GENETIC MAPS

Author
item Rohrer, Gary
item TROYER, D - KANSAS STATE UNIV.
item GOAD, D - KANSAS STATE UNIV.
item XIE, H - KANSAS STATE UNIV.
item Alexander, Leeson
item Beattie, Craig

Submitted to: Animal Genetics
Publication Type: Abstract Only
Publication Acceptance Date: 12/1/1994
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Direct in situ single copy PCR (DISC-PCR) procedures have many important uses for genomic research. We are using this technology to determine genomic coverage of a predominantly microsatellite based linkage map of the pig. We also use the results from DISC-PCR to orient and position linkage groups relative to cytogenetic benchmarks. Our first application of DISC-PCR was used to determine the proportion of pig chromosome 1 covered by a linkage group which contained 26 markers and spanned over 150 cM. Six microsatellites from this group were tested and physically assigned using the DISC-PCR technique. All assignments were comparable to those established by linkage analysis. Approximately 40% of pig chromosome 1 was covered by the linkage group, all markers were assigned to 1q, and orientation relative to the centromere was determined. Since all of the microsatellite markers assigned were sequenced from small genomic clones (<500 bp) the method allowed us to physically assign markers which could not have been conveniently done by currently available approaches. We are using the procedure in conjunction with FISH assignments of informative cosmids to determine and extend coverage of other linkage groups in the porcine genome. These results demonstrate the usefullness of a DISC-PCR strategy to rapidly produce comprehensive genome maps.