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A new bacteriophage P1-derived vector for the propagation of large human DNA fragments

Ioannou, Panayiotis A. and Amemiya, Chris T. and Garnes, Jeffrey and Kroisel, Peter M. and Shizuya, Hiroaki and Chen, Chira and Batzer, Mark and de Jong, Pieter J. (1994) A new bacteriophage P1-derived vector for the propagation of large human DNA fragments. Nature Genetics, 6 (1). pp. 84-89. ISSN 1061-4036. http://resolver.caltech.edu/CaltechAUTHORS:20150624-084912943

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Abstract

We have designed a P1 vector (pCYPAC−1) for the introduction of recombinant DNA into E. coli using electroporation procedures. The new cloning system, P1−derived arteficial chromosomes (PACs), was used to establish an initial 15,000 clone library with an average insert size of 130−150 kilobase pairs (kb). No chimaerism has been observed in 34 clones, by fluorescence in situ hybridization. Similarly, no insert instability has been observed after extended culturing, for 20 clones. We conclude that the PAC cloning system will be useful in the mapping and detailed analysis of complex genomes.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1038/ng0194-84 DOIArticle
http://www.nature.com/ng/journal/v6/n1/abs/ng0194-84.htmlPublisherArticle
Additional Information:© 1994 Nature Publishing Group. Received 28 September; accepted 18 October. P.A.I. and C. T.A contributed equally to this work. We thank Joel Jessee (BRL/Life Technologies, Inc.) Nat Sternberg for their help and contribution to this work. P.A.I. was partially supported by a grant from the Muscular Dystrophy Association, U.S.A., and by a Unesco/TWAS short term fellowship in the Human Genome Project. P.M.K. was supported in part by the FWF grant J0720-MED. Work performed at the Lawrence Livermore National Laboratory under the auspices of the U.S. Department of Energy under contract No. W-7405-ENG-48.
Funders:
Funding AgencyGrant Number
Muscular Dystrophy AssociationUNSPECIFIED
Unesco/TWASUNSPECIFIED
FWF Der WissenschaftsfondsJ0720-MED
Department of Energy (DOE)W-7405-ENG-48
Record Number:CaltechAUTHORS:20150624-084912943
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150624-084912943
Official Citation:loannou, P. A., Amemiya, C. T., Garnes, J., Kroisel, P. M., Shizuya, H., Chen, C., . . . de Jong, P. J. (1994). A new bacteriophage P1-derived vector for the propagation of large human DNA fragments. [10.1038/ng0194-84]. Nat Genet, 6(1), 84-89.
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ID Code:58573
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:24 Jun 2015 18:59
Last Modified:24 Jun 2015 18:59

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