Production of complex nucleic acid libraries using highly parallel in situ oligonucleotide synthesis View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2004-12

AUTHORS

Michele A Cleary, Kristopher Kilian, Yanqun Wang, Jeff Bradshaw, Guy Cavet, Wei Ge, Amit Kulkarni, Patrick J Paddison, Kenneth Chang, Nihar Sheth, Eric Leproust, Ernest M Coffey, Julja Burchard, W Richard McCombie, Peter Linsley, Gregory J Hannon

ABSTRACT

Generation of complex libraries of defined nucleic acid sequences can greatly aid the functional analysis of protein and gene function. Previously, such studies relied either on individually synthesized oligonucleotides or on cellular nucleic acids as the starting material. As each method has disadvantages, we have developed a rapid and cost-effective alternative for construction of small-fragment DNA libraries of defined sequences. This approach uses in situ microarray DNA synthesis for generation of complex oligonucleotide populations. These populations can be recovered and either used directly or immortalized by cloning. From a single microarray, a library containing thousands of unique sequences can be generated. As an example of the potential applications of this technology, we have tested the approach for the production of plasmids encoding short hairpin RNAs (shRNAs) targeting numerous human and mouse genes. We achieved high-fidelity clone retrieval with a uniform representation of intended library sequences. More... »

PAGES

241-248

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  • Journal

    TITLE

    Nature Methods

    ISSUE

    3

    VOLUME

    1

    From Grant

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nmeth724

    DOI

    http://dx.doi.org/10.1038/nmeth724

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1005983602

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/15782200


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