Abstract
Precise DNA manipulation is critical for molecular biotechnology. Restriction enzyme-based approaches are limited by their requirement of specific enzyme sites. Restriction-free cloning has greatly improved the flexibility and speed of precise DNA assembly. Most of these approaches focus on DNA assembly rather than gene removal. Here we present a polymerase chain reaction (PCR)-based cloning method that allows removal of multiple gene segments from plasmids without using restriction enzymes and thermostable ligase. We demonstrate simultaneous removal of three gene segments from a plasmid. This approach could be beneficial to DNA library construction, genetic and protein engineering, and synthetic biology.
Original language | English (US) |
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Pages (from-to) | 7-9 |
Number of pages | 3 |
Journal | Analytical Biochemistry |
Volume | 481 |
DOIs | |
State | Published - Jun 3 2015 |
Keywords
- Multiplex gene removal
- PCR
- Plasmids
- Restriction-free cloning
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology