Sean P Corum at 21:10, 25 June 2012

Sean P Corum — Mon, 25 Jun 2012 21:10:11 GMT

←Older revision		Revision as of 21:10, 25 June 2012
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	==Contact==		==Contact==
-	* [http://openwetware.org/wiki/User:Sean_P_Corum Sean P Corum] *'''SC 15:40, 23 June 2012 (EDT)''':	+	* [http://openwetware.org/wiki/User:Sean_P_Corum Sean P Corum]
		+
		+	==Digital Signature==
		+
		+	*'''SC 15:40, 23 June 2012 (EDT)''':

	or instead, [[Talk:{{PAGENAME}}\|discuss this protocol]].		or instead, [[Talk:{{PAGENAME}}\|discuss this protocol]].

Sean P Corum: New page: ==Overview== Transformation of chemically competent E. coli cells. ==Materials== * 20 μL frozen chemically competent cells (standard is JM109) * 1(3) μL high(low) concentration plasmid...

Sean P Corum — Sat, 23 Jun 2012 19:40:38 GMT

New page: ==Overview== Transformation of chemically competent E. coli cells. ==Materials== * 20 μL frozen chemically competent cells (standard is JM109) * 1(3) μL high(low) concentration plasmid...

New page

==Overview==
Transformation of chemically competent E. coli cells.

==Materials==

* 20 μL frozen chemically competent cells (standard is JM109)
* 1(3) μL high(low) concentration plasmid

==Procedure==
# Incubate competent cells on ice until liquid.
# Add plasmid. Quickly but gently mix and spin.
# Incubate on ice 10 min. In the meantime, heat water bath to 42 °C. (42 °C is standard. Some special strains may require a different temperature. For example, transformation into KL740 is done at 29 °C).
# Heat shock cells 20 s at 42 °C (or special temperature). Quickly return to ice. Incubate on ice an additional 2 min.
# Add 500 μL LB broth.
# Incubate in shaker 40 min 37 °C (or special temperature).
# Plate 30 μL cells onto an LB + antibiotic plate with a sterile pasture pipette. (Note, this is for high concentration plasmids. For low concentration plasmids, pellet cells by centrifugation, remove 500 μL supernatant, and re-suspend cells in the remaining 20 μL. Plate this 20 μL onto the LB plate.)
# Incubate overnight 37 °C (or special temperature).

==Notes==
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
#List troubleshooting tips here.
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
#Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.

==Contact==
* [http://openwetware.org/wiki/User:Sean_P_Corum Sean P Corum] *'''SC 15:40, 23 June 2012 (EDT)''':

or instead, [[Talk:{{PAGENAME}}|discuss this protocol]].



[[Category:Protocol]]

[[Category:Microbiology]]

[[Category:E. coli]]

[[Category:Transformation]]

Corum:Transformation - Revision history

Sean P Corum at 21:10, 25 June 2012

Sean P Corum: New page: ==Overview== Transformation of chemically competent E. coli cells. ==Materials== * 20 μL frozen chemically competent cells (standard is JM109) * 1(3) μL high(low) concentration plasmid...