Janet B. Matsen:Guide to Gibson Assembly: Difference between revisions
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(New page: * you can chose where the seam is if you use longer oligos * Dpn1 * RFP for backbone: don't screen red colonies!) |
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== 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 06:35, 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!
