| 摘要 |
<p>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.</p> |
