High-Efficiency Full-Length cDNA Cloning by Biotinylated CAP Trapper
We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryotic mRNA, followed by RNase I treatment to select full-length cDNA. The selection occurs by trapping the...
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| Veröffentlicht in: | Genomics (San Diego, Calif.) Calif.), 1996-11, Vol.37 (3), p.327-336 |
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| container_title | Genomics (San Diego, Calif.) |
| container_volume | 37 |
| creator | Carninci, Piero Kvam, Catrine Kitamura, Akiko Ohsumi, Tomoya Okazaki, Yasushi Itoh, Mitsuteru Kamiya, Mamoru Shibata, Kazuhiro Sasaki, Nobuya Izawa, Masaki Muramatsu, Masami Hayashizaki, Yoshihide Schneider, Claudio |
| description | We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryotic mRNA, followed by RNase I treatment to select full-length cDNA. The selection occurs by trapping the biotin residue at the cap sites using streptavidin-coated magnetic beads, thus eliminating incompletely synthesized cDNAs. When this method was used to construct a mouse brain full-length cDNA library, our evaluation showed that more than 95% of the total clones were of full length, and recombinant clones could be produced with high efficiency (1.2 × 107/10 μg starting mRNA). The analysis of 120 randomly picked clones indicates an unbiased representation of the starting mRNA population. |
| doi_str_mv | 10.1006/geno.1996.0567 |
| format | Article |
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The selection occurs by trapping the biotin residue at the cap sites using streptavidin-coated magnetic beads, thus eliminating incompletely synthesized cDNAs. When this method was used to construct a mouse brain full-length cDNA library, our evaluation showed that more than 95% of the total clones were of full length, and recombinant clones could be produced with high efficiency (1.2 × 107/10 μg starting mRNA). The analysis of 120 randomly picked clones indicates an unbiased representation of the starting mRNA population.</description><identifier>ISSN: 0888-7543</identifier><identifier>EISSN: 1089-8646</identifier><identifier>DOI: 10.1006/geno.1996.0567</identifier><identifier>PMID: 8938445</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Animals ; Bacterial Proteins ; Base Sequence ; Biological and medical sciences ; Biotin ; Brain Chemistry ; Chromatography, Affinity ; Cloning, Molecular - methods ; Diverse techniques ; DNA, Complementary - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Library ; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics ; Mice ; Molecular and cellular biology ; Molecular Sequence Data ; Peptide Elongation Factor 1 ; Peptide Elongation Factors - genetics ; Ribonuclease, Pancreatic - metabolism ; RNA Caps - chemistry ; RNA, Messenger - genetics ; Streptavidin</subject><ispartof>Genomics (San Diego, Calif.), 1996-11, Vol.37 (3), p.327-336</ispartof><rights>1996 Academic Press</rights><rights>1997 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c465t-c46e9884280bd15841ff83e2674434d3f86a9efc0152db114f98aad6d94bb8813</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/geno.1996.0567$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,45974</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2493508$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8938445$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carninci, Piero</creatorcontrib><creatorcontrib>Kvam, Catrine</creatorcontrib><creatorcontrib>Kitamura, Akiko</creatorcontrib><creatorcontrib>Ohsumi, Tomoya</creatorcontrib><creatorcontrib>Okazaki, Yasushi</creatorcontrib><creatorcontrib>Itoh, Mitsuteru</creatorcontrib><creatorcontrib>Kamiya, Mamoru</creatorcontrib><creatorcontrib>Shibata, Kazuhiro</creatorcontrib><creatorcontrib>Sasaki, Nobuya</creatorcontrib><creatorcontrib>Izawa, Masaki</creatorcontrib><creatorcontrib>Muramatsu, Masami</creatorcontrib><creatorcontrib>Hayashizaki, Yoshihide</creatorcontrib><creatorcontrib>Schneider, Claudio</creatorcontrib><title>High-Efficiency Full-Length cDNA Cloning by Biotinylated CAP Trapper</title><title>Genomics (San Diego, Calif.)</title><addtitle>Genomics</addtitle><description>We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryotic mRNA, followed by RNase I treatment to select full-length cDNA. 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Psychology</subject><subject>Gene Library</subject><subject>Glyceraldehyde-3-Phosphate Dehydrogenases - genetics</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><subject>Molecular Sequence Data</subject><subject>Peptide Elongation Factor 1</subject><subject>Peptide Elongation Factors - genetics</subject><subject>Ribonuclease, Pancreatic - metabolism</subject><subject>RNA Caps - chemistry</subject><subject>RNA, Messenger - genetics</subject><subject>Streptavidin</subject><issn>0888-7543</issn><issn>1089-8646</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkDFv2zAQRomiheOkXbsF0FB0k0tKJHUcHSeOCxhNhnQmKPLosJAlh5QL-N9Hig1vQZb7hnv34fAI-c7ojFEqf22w7WZMKTmjQlafyJRRUDlILj-TKQWAvBK8vCCXKf2jlKoSigmZwJCciym5XYXNc37nfbABW3vIlvumydfYbvrnzN7-mWeLpmtDu8nqQ3YTuj60h8b06LLF_DF7ima3w_iVfPGmSfjtlFfk7_LuabHK1w_3vxfzdW65FP04UQHwAmjtmADOvIcSC1lxXnJXepBGobeUicLVjHGvwBgnneJ1DcDKK_Lz2LuL3cseU6-3IVlsGtNit0-6AiFVofiHIJNUlFRWAzg7gjZ2KUX0ehfD1sSDZlSPfvXoV49-9eh3OLg-Ne_rLbozfhI67H-c9iZZ0_hoWhvSGSu4KgWFAYMjhoOu_wGjTm_-0YWItteuC-998ArjiJRE</recordid><startdate>19961101</startdate><enddate>19961101</enddate><creator>Carninci, Piero</creator><creator>Kvam, Catrine</creator><creator>Kitamura, Akiko</creator><creator>Ohsumi, Tomoya</creator><creator>Okazaki, Yasushi</creator><creator>Itoh, Mitsuteru</creator><creator>Kamiya, Mamoru</creator><creator>Shibata, Kazuhiro</creator><creator>Sasaki, Nobuya</creator><creator>Izawa, Masaki</creator><creator>Muramatsu, Masami</creator><creator>Hayashizaki, Yoshihide</creator><creator>Schneider, Claudio</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19961101</creationdate><title>High-Efficiency Full-Length cDNA Cloning by Biotinylated CAP Trapper</title><author>Carninci, Piero ; Kvam, Catrine ; Kitamura, Akiko ; Ohsumi, Tomoya ; Okazaki, Yasushi ; Itoh, Mitsuteru ; Kamiya, Mamoru ; Shibata, Kazuhiro ; Sasaki, Nobuya ; Izawa, Masaki ; Muramatsu, Masami ; Hayashizaki, Yoshihide ; Schneider, Claudio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c465t-c46e9884280bd15841ff83e2674434d3f86a9efc0152db114f98aad6d94bb8813</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Bacterial Proteins</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotin</topic><topic>Brain Chemistry</topic><topic>Chromatography, Affinity</topic><topic>Cloning, Molecular - methods</topic><topic>Diverse techniques</topic><topic>DNA, Complementary - genetics</topic><topic>Fundamental and applied biological sciences. 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| fulltext | fulltext |
| identifier | ISSN: 0888-7543 |
| ispartof | Genomics (San Diego, Calif.), 1996-11, Vol.37 (3), p.327-336 |
| issn | 0888-7543 1089-8646 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_78569294 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Bacterial Proteins Base Sequence Biological and medical sciences Biotin Brain Chemistry Chromatography, Affinity Cloning, Molecular - methods Diverse techniques DNA, Complementary - genetics Fundamental and applied biological sciences. Psychology Gene Library Glyceraldehyde-3-Phosphate Dehydrogenases - genetics Mice Molecular and cellular biology Molecular Sequence Data Peptide Elongation Factor 1 Peptide Elongation Factors - genetics Ribonuclease, Pancreatic - metabolism RNA Caps - chemistry RNA, Messenger - genetics Streptavidin |
| title | High-Efficiency Full-Length cDNA Cloning by Biotinylated CAP Trapper |
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