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Plant Cell, Tissue and Organ Culture (PCTOC) - 相似文献
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J.L. Corbett 《Animal Feed Science and Technology》1982,7(1):105-106
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J.K. Loosli 《Animal Feed Science and Technology》1981,6(2):209-210
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Two-dimensional screening of the Wageningen chicken BAC library 总被引:10,自引:0,他引:10
Richard P.M.A. Crooijmans Julia Vrebalov Rosilde J.M. Dijkhof Jan J. van der Poel Martien A.M. Groenen 《Mammalian genome》2000,11(5):360-363
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.
Received: 26 October 1999 / Accepted: 6 January 2000 相似文献
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