Chapter title |
Simultaneous Removal of Multiple DNA Segments by Polymerase Chain Reactions.
|
---|---|
Chapter number | 15 |
Book title |
Synthetic DNA
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6343-0_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6341-6, 978-1-4939-6343-0
|
Authors |
Vishnu Krishnamurthy, Kai Zhang Ph.D., Kai Zhang |
Editors |
Randall A. Hughes |
Abstract |
Precise DNA manipulation is a key enabling technology for synthetic biology. Approaches based on restriction digestion are often limited by the presence of certain restriction enzyme recognition sites. Recent development of restriction-free cloning approaches has greatly enhanced the flexibility and speed of molecular cloning. Most restriction-free cloning methods focus on DNA assembly. Much less work has been dedicated towards DNA removal. Here we introduce a protocol that allows simultaneous removal of multiple DNA segments from a plasmid using polymerase chain reactions (PCR). Our approach will be beneficial to applications in multiple sites mutagenesis, DNA library construction, genetic and protein engineering, and synthetic biology. |
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