Janet B. Matsen:Guide to Gibson Assembly
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| + | Back to [[User:Janet B. Matsen|Janet]] | ||
| + | |||
| + | == Intro == | ||
| + | * 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 | ||
| + | ** Transform | ||
| + | *** Electroporation is usually used to provide higher yield. | ||
| + | |||
| + | |||
| + | == Examples == | ||
| + | |||
| + | |||
* you can chose where the seam is if you use longer oligos | * you can chose where the seam is if you use longer oligos | ||
* Dpn1 | * Dpn1 | ||
* RFP for backbone: don't screen red colonies! | * RFP for backbone: don't screen red colonies! | ||
Revision as of 09:35, 9 October 2012
Back to Janet
Intro
- 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
- Transform
- Electroporation is usually used to provide higher yield.
Examples
- you can chose where the seam is if you use longer oligos
- Dpn1
- RFP for backbone: don't screen red colonies!
