Two-dimensional screening of the Wageningen chicken BAC library (original) (raw)

Abstract.

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 _Hin_dIII-digested high-molecular-weight (HMW) DNA of a female White Leghorn chicken into the _Hin_dIII site of the vector pECBAC1. Several modifications of standard protocols were necessary to clone efficiently large partial _Hin_dIII 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 _Not_I 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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Authors and Affiliations

  1. Animal 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
  2. Department of Horticultural Sciences and Crop Biotechnology centre, Texas A&M University, College Station, TX, USA, , , , , , US
    Julia Vrebalov

Authors

  1. Richard P.M.A. Crooijmans
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  2. Julia Vrebalov
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  3. Rosilde J.M. Dijkhof
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  4. Jan J. van der Poel
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  5. Martien A.M. Groenen
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Received: 26 October 1999 / Accepted: 6 January 2000

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Crooijmans, 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

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