Janet B. Matsen:Guide to Gibson Assembly
Back to Janet
- What is it?
- How does it differ from other cloning?
- When should I use it?
- Steps (concise)
- Design oligos to yield 40 - 100 bp overlapping linear DNA segments
- Purify (usually gel) the PCR products (or digest)
- Use Gibson Assembly Mix
- Electroporation is usually used to provide higher yield.
- you can chose where the seam is if you use longer oligos
- RFP for backbone: don't screen red colonies!