Summary
A universally primed polymerase chain reaction was developed to amplify DNA dissected from GTG-banded human chromosomes. The amplification products are cloned into plasmid vectors, which allow the rapid characterization of recombinant clones. Starting from 20–40 chromosome fragments, several thousand independent clones detecting single-copy sequences can be obtained. Although these libraries comprise only a few percent of the dissected DNA, they provide narrowly spaced anchor clones for the molecular characterization of chromosome bands and the identification of gene sequences. Here we describe the construction and characterization of DNA libraries for the Langer-Giedion syndrome chromosome region (LGCR, 8q23–24.1), Wilms tumor chromosome region 1 (WT1, 11p13), Prader-Willi syndrome/Angelman syndrome chromosome region (PWCR/ANCR, 15q11.2–12), meningioma chromosome region (MGCR, 22q12–13), and fragile X chromosome region (FRAXA, Xq27.3).
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References
Bishop DT, Williamson JA, Skolnick MH (1983) A model for restriction fragment length distributions. Am J Hum Genet 35:795–815
Brown SDM, Greenfield AJ (1987) A model to describe the size distribution of mammalian genomic fragments recovered by microcloning. Gene 55:327–332
Buiting K, Neumann M, Lüdecke HJ, Senger G, Claussen U, Antich J, Passarge E, Horsthemke B (1990) Microdissection of potential gene sequences from the Prader-Willi syndrome chromosome region. Genomics (in press)
Deininger PL, Jolly DJ, Rubin CM, Friedmann T, Schmid CW (1981) Base sequence studies of 300 nucleotide renatured repeated human DNA clones. J Mol Biol 151:17–33
Donlon TA, Lalande M, Wyman A, Bruns G, Latt SA (1986) Isolation of molecular probes associated with the chromosome 15 instability in the Prader-Willi syndrome. Proc Natl Acad Sci USA 83:4408–4412 (correction 83:6964)
Edström JE, Kaiser R, Röhme D (1987) Microcloning of mammalian metaphase chromosomes. Methods Enzymol 151:503–516
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Horsthemke B, Greger V, Barnert HJ, Höpping W, Passarge E (1987) Detection of submicroscopic deletions and a DNA polymorphism at the retinoblastoma locus. Hum Genet 76:257–261
Kaiser R, Weber J, Grzeschik K-H, Edström JE, Driesel A, Zengerling S, Buchwald M, Tsui LC, Olek K (1987) Microdissection and microcloning of the long arm of human chromosome 7. Mol Biol Rep 12:3–6
Kunkel TA, Schaaper RM, Loeb LA (1983) Depurination induced infidelity of deoxyribonucleic acid synthesis with purified deoxyribonucleic acid replication proteins in vitro. Biochemistry 22:2378–2384
Langley KE, Villarejo MR, Fowler AV, Zamenhof PJ, Zabin I (1975) Molecular basis of β-galactosidase-α complementation. Proc Natl Acad Sci USA 72:1254–1257
Lüdecke HJ, Senger G, Claussen U, Horsthemke B (1989) Cloning of defined regions of the human genome by microdissection of banded chromsomes and enzymatic amplification. Nature 338:348–350
Messing J, Gronenborn B (1978) The filamentous phage M13 as a carrier DNA for operon fusions in vitro. In: Denhardt DT, Dressler D, Ray DS (eds) The single-stranded DNA phages. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 449–453
Pfeiffer BH, Zimmermann SB (1983) Polymer-stimulated ligation: enhanced blunt- or cohesive-end ligation of DNA or deoxyribo-oligonucleotides by T4 DNA ligase in polymer solutions. Nucleic Acids Res 11:7853–7871
Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230:1350–1354
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Sanger F, Nicklen S, Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Senger G, Lüdecke H-J, Horsthemke B, Claussen U (1990) Microdissection of banded human chromosomes. Hum Genet 84:507–511
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Lüdecke, HJ., Senger, G., Claussen, U. et al. Construction and characterization of band-specific DNA libraries. Hum Genet 84, 512–516 (1990). https://doi.org/10.1007/BF00210800
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DOI: https://doi.org/10.1007/BF00210800