Team:UC Davis/Notebook/Protocols
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<br>•Incubate at 37 °C for 1 hour. | <br>•Incubate at 37 °C for 1 hour. | ||
<br>•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic. | <br>•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic. | ||
- | <br>•Incubate overnight at 37 °C. | + | <br>•Incubate overnight at 37 °C<html><span onmousedown="(function (srcs,cfg) { var cbcount = 1; var callback = function () { -- cbcount; if (cbcount === 0) { BrowserPonies.setBaseUrl(cfg.baseurl); if (!BrowserPoniesBaseConfig.loaded) { BrowserPonies.loadConfig(BrowserPoniesBaseConfig); BrowserPoniesBaseConfig.loaded = true; } BrowserPonies.loadConfig(cfg); if (!BrowserPonies.running()) BrowserPonies.start(); } }; if (typeof(BrowserPoniesConfig) === "undefined") { window.BrowserPoniesConfig = {}; } if (typeof(BrowserPoniesBaseConfig) === "undefined") { ++ cbcount; BrowserPoniesConfig.onbasecfg = callback; } if (typeof(BrowserPonies) === "undefined") { ++ cbcount; BrowserPoniesConfig.oninit = callback; } var node = (document.body || document.documentElement || document.getElementsByTagName('head')[0]); for (var id in srcs) { if (document.getElementById(id)) continue; if (node) { var s = document.createElement('script'); s.type = 'text/javascript'; s.id = id; s.src = srcs[id]; node.appendChild(s); } else { document.write('\u003cscript type="text/javscript" src="'+ srcs[id]+'" id="'+id+'"\u003e\u003c/script\u003e'); } } callback(); |
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Revision as of 23:07, 31 July 2012
Protocols
•Add 20ul sterile H2O to desired well in distribution plate.
•Incubate at room temperature for ~10 minutes.
•Transfer resuspension to microcentrifuge tube.
Materials
•Competent Cells•DNA template
•800 ul LB
•LB+Antibiotic Plates
Procedure
•Thaw competent cells on ice.•Transfer 50 uL of competent cells to chilled falcon tubes.
•Add 1 uL of template to cells (2.5 uL if dilute).
•Incubate on ice for 30 minutes.
•Heat schock in 42 °C water bath for 90 seconds.
•Immediately place back onto ice for 2 minutes.
•Add 800 uL of LB to each tube.
•Incubate at 37 °C for 1 hour.
•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic.
•Incubate overnight at 37 °C.