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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
1997-3-27
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pubmed:abstractText |
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 x 10(7)/10 micrograms starting mRNA). The analysis of 120 randomly picked clones indicates an unbiased representation of the starting mRNA population.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Biotin,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Glyceraldehyde-3-Phosphate...,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Elongation Factor 1,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Elongation Factors,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger,
http://linkedlifedata.com/resource/pubmed/chemical/RNA Caps,
http://linkedlifedata.com/resource/pubmed/chemical/Ribonuclease, Pancreatic,
http://linkedlifedata.com/resource/pubmed/chemical/Streptavidin
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0888-7543
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pubmed:author |
pubmed-author:CarninciPP,
pubmed-author:HayashizakiYY,
pubmed-author:ItohMM,
pubmed-author:IzawaMM,
pubmed-author:KamiyaMM,
pubmed-author:KitamuraAA,
pubmed-author:KvanLL,
pubmed-author:MuramatsuMM,
pubmed-author:OhsumiTT,
pubmed-author:OkazakiYY,
pubmed-author:SasakiNN,
pubmed-author:SchneiderCC,
pubmed-author:ShibataKK
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
37
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
327-36
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:8938445-Animals,
pubmed-meshheading:8938445-Bacterial Proteins,
pubmed-meshheading:8938445-Base Sequence,
pubmed-meshheading:8938445-Biotin,
pubmed-meshheading:8938445-Brain Chemistry,
pubmed-meshheading:8938445-Chromatography, Affinity,
pubmed-meshheading:8938445-Cloning, Molecular,
pubmed-meshheading:8938445-DNA, Complementary,
pubmed-meshheading:8938445-Gene Library,
pubmed-meshheading:8938445-Glyceraldehyde-3-Phosphate Dehydrogenases,
pubmed-meshheading:8938445-Mice,
pubmed-meshheading:8938445-Molecular Sequence Data,
pubmed-meshheading:8938445-Peptide Elongation Factor 1,
pubmed-meshheading:8938445-Peptide Elongation Factors,
pubmed-meshheading:8938445-RNA, Messenger,
pubmed-meshheading:8938445-RNA Caps,
pubmed-meshheading:8938445-Ribonuclease, Pancreatic,
pubmed-meshheading:8938445-Streptavidin
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pubmed:year |
1996
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pubmed:articleTitle |
High-efficiency full-length cDNA cloning by biotinylated CAP trapper.
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pubmed:affiliation |
Genome Science Laboratory, Tsukuba Life Science Center, Ibaraki, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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