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{"title"=>"A colony multiplex quantitative PCR-based 3S3DBC method and variations of it for screening DNA libraries", "type"=>"journal", "authors"=>[{"first_name"=>"Yang", "last_name"=>"An", "scopus_author_id"=>"56528862300"}, {"first_name"=>"Atsushi", "last_name"=>"Toyoda", "scopus_author_id"=>"55488478100"}, {"first_name"=>"Chen", "last_name"=>"Zhao", "scopus_author_id"=>"56506018800"}, {"first_name"=>"Asao", "last_name"=>"Fujiyama", "scopus_author_id"=>"7003611650"}, {"first_name"=>"Kiyokazu", "last_name"=>"Agata", "scopus_author_id"=>"7005905990"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84922341489", "pui"=>"602005550", "pmid"=>"25646755", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0116997", "sgr"=>"84922341489"}, "id"=>"b5d2f2cd-3d02-3a62-b143-effb8f52a3fc", "abstract"=>"A DNA library is a collection of DNA fragments cloned into vectors and stored individually in host cells, and is a valuable resource for molecular cloning, gene physical mapping, and genome sequencing projects. To take the best advantage of a DNA library, a good screening method is needed. After describing pooling strategies and issues that should be considered in DNA library screening, here we report an efficient colony multiplex quantitative PCR-based 3-step, 3-dimension, and binary-code (3S3DBC) method we used to screen genes from a planarian genomic DNA fosmid library. This method requires only 3 rounds of PCR reactions and only around 6 hours to distinguish one or more desired clones from a large DNA library. According to the particular situations in different research labs, this method can be further modified and simplified to suit their requirements.", "link"=>"http://www.mendeley.com/research/colony-multiplex-quantitative-pcrbased-3s3dbc-method-variations-it-screening-dna-libraries", "reader_count"=>5, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Student > Master"=>1, "Professor"=>1, "Student > Ph. D. Student"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Student > Master"=>1, "Professor"=>1, "Student > Ph. D. Student"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>4}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/1879666"], "description"=>"<p>This figure was snipped from our results obtained using ABI qPCR analysis software SDS 2.4 standalone, and shows representative results of disassociation curves generated from the first step of 3S3DBC DNA screening. Positive DNA template (10 ng of <i>Dugesia japonica</i> genomic DNA) was used to detect the qPCR specificity of the respective primer set of DjPiwiB (a) and DJF-033N19 (b). Each primer set could also generate the specific qPCR product after qPCR in a real library super pool (c and d). No nonspecific amplification was detected in the negative super pool or in the H₂O negative template control, using either DjPiwiB or DJF-033N19 multiplex primer sets (e and f). The expected qPCR disassociation curve (double peak shows two qPCR products) was detected in the positive library super pool using DjPiwiB and DJF-033N19 multiplex primer sets (g), and the dissociation temperature values of the two peaks corresponded to the same values obtained in the positive template control and positive super pool results. All figures from (a) to (g) were merged to produce (h).</p>", "links"=>[], "tags"=>["pcr", "DNA library", "genome sequencing projects", "method", "Screening DNA Libraries", "DNA library screening", "planarian genomic DNA fosmid library", "3DBC"], "article_id"=>1300702, "categories"=>["Biological Sciences"], "users"=>["Yang An", "Atsushi Toyoda", "Chen Zhao", "Asao Fujiyama", "Kiyokazu Agata"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0116997.g002", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Representative_results_of_qPCR_disassociation_curves_during_screening_/1300702", "title"=>"Representative results of qPCR disassociation curves during screening.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-03 03:23:03"}
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• {"files"=>["https://ndownloader.figshare.com/files/1879717", "https://ndownloader.figshare.com/files/1879718", "https://ndownloader.figshare.com/files/1879719", "https://ndownloader.figshare.com/files/1879721", "https://ndownloader.figshare.com/files/1879722", "https://ndownloader.figshare.com/files/1879724", "https://ndownloader.figshare.com/files/1879725", "https://ndownloader.figshare.com/files/1879726", "https://ndownloader.figshare.com/files/1879727", "https://ndownloader.figshare.com/files/1879729", "https://ndownloader.figshare.com/files/1879730", "https://ndownloader.figshare.com/files/1879731"], "description"=>"<div><p>A DNA library is a collection of DNA fragments cloned into vectors and stored individually in host cells, and is a valuable resource for molecular cloning, gene physical mapping, and genome sequencing projects. To take the best advantage of a DNA library, a good screening method is needed. After describing pooling strategies and issues that should be considered in DNA library screening, here we report an efficient colony multiplex quantitative PCR-based 3-step, 3-dimension, and binary-code (3S3DBC) method we used to screen genes from a planarian genomic DNA fosmid library. This method requires only 3 rounds of PCR reactions and only around 6 hours to distinguish one or more desired clones from a large DNA library. According to the particular situations in different research labs, this method can be further modified and simplified to suit their requirements.</p></div>", "links"=>[], "tags"=>["pcr", "DNA library", "genome sequencing projects", "method", "Screening DNA Libraries", "DNA library screening", "planarian genomic DNA fosmid library", "3DBC"], "article_id"=>1300743, "categories"=>["Biological Sciences"], "users"=>["Yang An", "Atsushi Toyoda", "Chen Zhao", "Asao Fujiyama", "Kiyokazu Agata"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0116997.s001", "https://dx.doi.org/10.1371/journal.pone.0116997.s002", "https://dx.doi.org/10.1371/journal.pone.0116997.s003", "https://dx.doi.org/10.1371/journal.pone.0116997.s004", "https://dx.doi.org/10.1371/journal.pone.0116997.s005", "https://dx.doi.org/10.1371/journal.pone.0116997.s006", "https://dx.doi.org/10.1371/journal.pone.0116997.s007", "https://dx.doi.org/10.1371/journal.pone.0116997.s008", "https://dx.doi.org/10.1371/journal.pone.0116997.s009", "https://dx.doi.org/10.1371/journal.pone.0116997.s010", "https://dx.doi.org/10.1371/journal.pone.0116997.s011", "https://dx.doi.org/10.1371/journal.pone.0116997.s012"], "stats"=>{"downloads"=>1, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_Colony_Multiplex_Quantitative_PCR_Based_3S3DBC_Method_and_Variations_of_It_for_Screening_DNA_Libraries_/1300743", "title"=>"A Colony Multiplex Quantitative PCR-Based 3S3DBC Method and Variations of It for Screening DNA Libraries", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-02-03 03:23:03"}

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• {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
• {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
• {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2017", "month"=>"1"}
• {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
• {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
• {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"4"}
• {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"5"}
• {"unique-ip"=>"8", "full-text"=>"6", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2017", "month"=>"6"}
• {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"7"}
• {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"8"}
• {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
• {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}
• {"unique-ip"=>"4", "full-text"=>"7", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"11"}
• {"unique-ip"=>"3", "full-text"=>"4", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
• {"unique-ip"=>"4", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
• {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
• {"unique-ip"=>"7", "full-text"=>"6", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"1"}
• {"unique-ip"=>"6", "full-text"=>"8", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
• {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
• {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"4"}
• {"unique-ip"=>"10", "full-text"=>"27", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"5"}
• {"unique-ip"=>"11", "full-text"=>"3", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"12", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
• {"unique-ip"=>"7", "full-text"=>"2", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
• {"unique-ip"=>"5", "full-text"=>"7", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
• {"unique-ip"=>"11", "full-text"=>"12", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
• {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
• {"unique-ip"=>"7", "full-text"=>"11", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
• {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
• {"unique-ip"=>"11", "full-text"=>"14", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
• {"unique-ip"=>"9", "full-text"=>"12", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
• {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
• {"unique-ip"=>"10", "full-text"=>"8", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
• {"unique-ip"=>"20", "full-text"=>"13", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"13", "cited-by"=>"1", "year"=>"2019", "month"=>"10"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}

F1000Prime