We have constructed a Bacterial Artificial Chromosome (BAC) library that provides 5.5-fold redundant coverage of the chicken genome. The library was made by cloning partial HindIII-digested high-molecular-weight (HMW) DNA of a female White Leghorn chicken into the HindIII site of the vector pECBAC1. Several modifications of standard protocols were necessary to clone efficiently large partial HindIII DNA fragments. The library consists of 49,920 clones arranged in 130 384-well plates. An average insert size of 134 kb was estimated from the analysis of 152 randomly selected BAC clones. The average number of NotI restriction sites per clone was 0.77. After individual growth, DNA was isolated of the pooled clones of each 384-well plate, and subsequently DNA of each plate was isolated from the individual row and column pools. Screening of the Wageningen chicken BAC library was performed by two-dimensional PCR with 125 microsatellite markers. For 124 markers at least one BAC clone was obtained. FISH experiments of 108 BAC clones revealed chimerism in less than 1%. The number of different BAC clones per marker present in the BAC library was examined for 35 markers which resulted in a total of 167 different BAC clones. Per marker the number of BAC clones varied from 1 to 11, with an average of 4.77. The chicken BAC library constitutes an invaluable tool for positional cloning and for comparative mapping studies.
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Similar content being viewed by others Explore related subjectsDiscover the latest articles and news from researchers in related subjects, suggested using machine learning. Author information Authors and AffiliationsAnimal Breeding and Genetics group, Wageningen Institute of Animal Sciences, Wageningen Agricultural University, Postbox 338, 6700 AH, Wageningen, The Netherlands, , , , , , NL
Richard P.M.A. Crooijmans, Rosilde J.M. Dijkhof, Jan J. van der Poel & Martien A.M. Groenen
Department of Horticultural Sciences and Crop Biotechnology centre, Texas A&M University, College Station, TX, USA, , , , , , US
Julia Vrebalov
Received: 26 October 1999 / Accepted: 6 January 2000
About this article Cite this articleCrooijmans, R., Vrebalov, J., Dijkhof, R. et al. Two-dimensional screening of the Wageningen chicken BAC library. 11, 360–363 (2000). https://doi.org/10.1007/s003350010068
Issue Date: May 2000
DOI: https://doi.org/10.1007/s003350010068
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