Electroporation

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Current revision (13:40, 23 June 2009) (view source)
(Specific Protocols)
 
(5 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:E.Coli Protocol]]
+
[[Category:Protocol]]
 +
[[Category:In vivo]]
 +
[[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.
Personal tools