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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
1996-6-13
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pubmed:databankReference |
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007128,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007129,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007130,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007131,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007132,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007133,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007134,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007135,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007136,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007137,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007138,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007139,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007140,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007141,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007142,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007143,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007144,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007145,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007146,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007147,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007148,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007149,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007150,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007151,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007152,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007153,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/AF007154
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pubmed:abstractText |
The efficiency of shotgun DNA sequencing depends to a great extent on the quality of the random-subclone libraries used. We here describe a novel "double adaptor" strategy for efficient construction of high-quality shotgun libraries. In this method, randomly sheared and end-repaired fragments are ligated to oligonucleotide adaptors creating 12-base overhangs. Nonphosphorylated oligonucleotides are used, which prevents formation of adaptor dimers and ensures efficient ligation of insert to adaptor. The vector is prepared from a modified M13 vector, by KpnI/PstI digestion followed by ligation to oligonucleotides with ends complementary to the overhangs created in the digest. These adaptors create 5'-overhangs complementary to those on the inserts. Following annealing of insert to vector, the DNA is directly used for transformation without a ligation step. This protocol is robust and shows three- to fivefold higher yield of clones compared to previous protocols. No chimeric clones can be detected and the background of clones without an insert is <1%. The procedure is rapid and shows potential for automation.
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pubmed:grant |
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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 |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0003-2697
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
236
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
107-13
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
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pubmed:year |
1996
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pubmed:articleTitle |
A "double adaptor" method for improved shotgun library construction.
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pubmed:affiliation |
Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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