Polymorphic Markers for Chromosome Band 11e2
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AbstractOur research decribes a strategy for producing a high density set of PCR-base microsatellite markerpolymorphic in L5178Y, the cell line used in the mouse lymphoma assay (MLA). These polymorphic marketwo detailed long range restriction maps in the vicinity of the thymidine kinase (tk-1) gene, with tdefined distances from each other and the tk-1 gene. The first, lower resolution map will define thethe tk-1 gene, and will contain a PCR-based polymorphic marker approximately every 1 Mb. Multiplex Pbe identified so that mutants can be assayed through the entire 10 MB regon in a single reaction, atThe second, higher resolution map will contain apporoximately 10 PCR-based polymorphic markers, posikb, in the 1 Mb region in the immediate vicinity of the tk-1 gene. Multiplex PCR conditions will alssecond set of markers so that mutants can be analyzed in this 1 Mb region at a resolution of 100 kb.these polymorphic markers, we will construct a genomic library of mouse chromosome band 11E2, accordfollowing strategy: Band 11E2 will be microdissected form normal metaphase spreads, PCR amplifies usoligonucleotide primer, and cloned. The library will be screened with a microsatellite repeat probe,(CA)n repeats will be sequenced. PCR primer sets flanking the repeats will be designed and synthesizbe assayed for heterozygosily. Polymorphis (CA)n repeats will be identifies by the production of two
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