Electroporation
From OpenWetWare
(Difference between revisions)
Current revision (14:40, 23 June 2009) (view source) (→Specific Protocols) |
|||
| (3 intermediate revisions not shown.) | |||
| Line 1: | Line 1: | ||
| + | {{back to protocols}} | ||
==Specific Protocols== | ==Specific Protocols== | ||
*[[Knight:Electroporation]] | *[[Knight:Electroporation]] | ||
| - | + | *[[Endy:High-efficiency electroporation]] | |
| - | + | *[[Richard Lab:Electroporation of E. coli]] | |
==Notes== | ==Notes== | ||
| - | + | *If you are in a hurry and your selection marker is ampicillin and you're sure to have enough plasmid (e.g. not after difficult clonings), you can go ahead and plate immediately because ampicillin takes a while to be pumped into cells at a high enough concentration to have an effect. This is called [http://www.neb.com/nebecomm/products/protocol120.asp 5-minute transformation] by New England Biolabs and if you follow the link you'll get a nice complete protocol for it (10% efficiency compared to full protocol) | |
| - | *If you are in a hurry and your selection marker is ampicillin, you can go ahead and plate immediately because ampicillin takes a while to be pumped into cells at a high enough concentration to have an effect. | + | |
| - | + | ||
*The [[Endy Lab]] is trying to use a standard positive control to better compare (and hopefully improve) the transformation efficiencies in the lab, you can check it out [[Endy:Standard transformation positive control|here]]. | *The [[Endy Lab]] is trying to use a standard positive control to better compare (and hopefully improve) the transformation efficiencies in the lab, you can check it out [[Endy:Standard transformation positive control|here]]. | ||
| - | |||
| - | |||
[[Category:Protocol]] | [[Category:Protocol]] | ||
| + | [[Category:In vivo]] | ||
[[Category:Escherichia coli]] | [[Category:Escherichia coli]] | ||
Current revision
| back to protocols | ||
Specific Protocols
Notes
- If you are in a hurry and your selection marker is ampicillin and you're sure to have enough plasmid (e.g. not after difficult clonings), you can go ahead and plate immediately because ampicillin takes a while to be pumped into cells at a high enough concentration to have an effect. This is called 5-minute transformation by New England Biolabs and if you follow the link you'll get a nice complete protocol for it (10% efficiency compared to full protocol)
- The Endy Lab is trying to use a standard positive control to better compare (and hopefully improve) the transformation efficiencies in the lab, you can check it out here.
