Rapid mapping of markers applying vectorette technology to YAC fragmentation allows easy assembly of a high-density STS bacterial clone contig spanning the markers D6S1260-D6S1918

Springer Science and Business Media LLC - Tập 9 - Trang 220-225 - 1998
Jeremy D. Shearman1,2, Jennifer J. Pointon1, Alison T. Merryweather-Clarke1, Caroline Stone1, Sharon W. Horsley1, Lyndal Kearney1, William M. Rosenberg2, Kathryn J. H. Robson1
1MRC Molecular Haematology Unit, Institute of Molecular Medicine, Headington, UK
2Nuffield Department of Medicine, Institute of Molecular Medicine, Headington, UK

Tóm tắt

We have generated a detailed physical map of the 6p21.3/p22.1 boundary, using a combination of yeast artificial chromosome (YAC) fragmentation and high-resolution sequence tagged site (STS) content mapping. YACs from the CEPH, St. Louis, and ICRF libraries have been used to construct a 4.5-Mb contig spanning the markers D6S306 to D6S1571. YAC insert sizes were determined by pulsed field gel electrophoresis (PFGE). Chimerism of YACs was determined by fluorescent in situ hybridization (FISH), and their integrity was determined by fingerprinting with Alu-PCR. We have identified 10 new CA repeat loci in this region as well as over 50 novel STSs, several tRNA genes, a new histone H2B gene and the phospholipase D gene. Using these new markers, we have rapidly generated a bacterial clone contig of over 250 kb, spanning the markers D6S1260 to D6S1918 (WI-3111) with STSs spaced on average every 6 kb.

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