| 摘要 |
A method for the automated molecular cloning and amplification of closed circular clones in vitro in a cell-free system using a pair of particularly designed cloning primers in a ligation-during-amplification (LDA) process, in which the fully extended DNA strands containing the donor sequence are ligated by DNA ligase and then used as templates for subsequent amplification. By designing particular cloning primers, the present invention can be used to obtain amplified closed circular nucleic acid clones with inverted or non-inverted insertions or substitutions with a donor nucleic acid fragment, or deletions of a fragment of the closed circular nucleic acid. |
