Construction of chromosome-specific DNA libraries covering the whole genome of field bean (Vicia faba L.)

Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology
J MacasJ Dolezel

Abstract

Recombinant DNA libraries were constructed for seven chromosome types isolated from two translocation lines of field bean (Vicia faba L.) with reconstructed karyotypes. The chromosomes were selected so that the set of libraries covers the whole V. faba genome more than once. Individual chromosome types were highly purified by flow sorting, and their DNA was amplified by degenerate oligonucleotide-primed (DOP) polymerase chain reaction (PCR) and cloned into a plasmid vector. The choice of restriction site present in PCR primer and refinement of cloning protocol resulted in high cloning efficiency and allowed generation of libraries consisting of about 10(5) clones from 250 or 1000 sorted chromosomes. The insert size ranged between 50 and 2200 bp and the mean length estimated in individual libraries varied between 310 and 487 bp. Hybridization of cloned fragments with labelled genomic DNA showed that about 60% of inserts represented unique or low-copy sequences. The suitability of the libraries for genome mapping was demonstrated by isolation of clones containing microsatellite motifs.

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Citations

Aug 17, 2012·Functional & Integrative Genomics·Jaroslav DoleželHana Simková
Aug 15, 2014·The Journal of Heredity·Emily RoseAdam G Jones
Jan 11, 2014·Biotechnology Advances·Jaroslav DoleželHana Simková
Sep 24, 2004·Hereditas·Lijia LiYunchun Song
Mar 3, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·David W Galbraith
Nov 11, 2003·Genome Génome / Conseil National De Recherches Canada·M KubalákováJ Dolezel
Sep 5, 2008·Current Protocols in Cytometry·J DolezelS Lucretti
Feb 23, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Jaroslav DoleželDebora Giorgi

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