NUCLEOTIDE CLONING METHODS
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United States of America Patent
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Issued Date -
Dec 11, 2014
app pub date -
Jan 11, 2013
filing date -
Jan 13, 2012
priority date (Note) -
Abandoned
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Abstract
Methods of cloning insert sequences into cloning vectors with high efficiency and in the correct orientation are described. In one aspect, the invention features a method of producing a plasmid comprising an insert fragment and a vector fragment in a predetermined orientation. In some embodiments, the method includes cleaving a first nucleotide sequence at a plurality of sites with a first restriction enzyme to generate a first population of nucleotide fragments, the first population of nucleotide fragments comprising insert fragments and non-insert fragments, the insert fragments comprising a non-palindromic overhang at a 5′ end, at a 3′ end, or at both.
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