Transformation of Plasmid DNA into E. coli Using the Heat Shock Method
Alexandrine Froger, James E. Hall
Department of Physiology and Biophysics, University of California, Irvine
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0:04 Introduction4 0:21 Subscription Lock21 0:38 Preparing for Heat Transformation38 1:14 Introducing plasmid DNA into bacteria by heat shock74 3:05 Plating transformed bacteria on LB agar plate185 5:59 Assessing the transformation efficiency359 6:42 Conclusion402
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Transformation of plasmid DNA into E. coli using the heat shock method is a basic technique of molecular biology. It consists of inserting a foreign plasmid or ligation product into bacteria. This video protocol describes the traditional method of transformation using commercially available chemically competent bacteria from Genlantis. After a short incubation in ice, a mixture of chemically competent bacteria and DNA is placed at 42°C for 45 seconds (heat shock) and then placed back in ice. SOC media is added and the transformed cells are incubated at 37°C for 30 min with agitation. To be assured of isolating colonies irrespective of transformation efficiency, two quantities of transformed bacteria are plated. This traditional protocol can be used successfully to transform most commercially available competent bacteria. The turbocells from Genlantis can also be used in a novel 3-minute transformation protocol, described in the instruction manual.
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Froger A, Hall JE.. (2007). Transformation of Plasmid DNA into E. coli Using the Heat Shock Method. JoVE. 6. http://www.jove.com/index/details.stp?id=253, doi: 10.3791/253
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Allowed tags: i, b, u, sup, sub 
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| | 02/01/2008 7:57:17 PM
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Gbeebani responded with a statement of type: Neutral
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I have never seen or heard anyone use beads to create a spread plate. Is that a new technique that has not reached my university yet, or is it an outdated technique that scientists don't use anymore?
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| | 12/24/2008 9:41:51 AM
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ali responded with a statement of type: Agree
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hi my neam isalirezababazade . iam student in unversity tehran . what do you teransfer factors F(negativ)in the e.coli?why transfe geneboth factor? |
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| | 02/01/2008 8:11:46 PM
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labrat responded with a statement of type: Neutral
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People use it all the time. Ask in the labs around, I am sure you will find people who use it.
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| | 03/09/2008 8:45:48 PM
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Anonymous responded with a statement of type: Agree
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I haven't seen it either so you're not alone. I'm wondering if it isn't easily to use a spreader in terms of cleanup and reducing contamination? |
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| | 09/09/2008 9:04:39 PM
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gulozgur responded with a statement of type: Neutral
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Copacabana method for spreading E. coli and yeast colonies
Mark T. Worthington, Roger Qi Luo, and Jared Pelo
BioTechniques Vol. 30, No. 4: pp 738-741 (Apr 2001) |
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| | 09/09/2008 4:21:53 PM
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Anonymous responded with a statement of type: Question
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why don't the flame is open? Don't you consider the contamination risk, or do you think that it is not necessary?? |
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| | 09/09/2008 4:22:19 PM
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zerrin responded with a statement of type: Question
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why don't the flame is open? Don't you consider the contamination risk, or do you think that it is not necessary?? |
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| | 09/16/2008 8:57:16 PM
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lab responded with a statement of type: Neutral
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Please look more carefully. The bunsen burner is clearly on and the blue flame is clearly visible in the video. It is on at all stages which are succeptible to contamination. |
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| | 09/09/2008 4:25:15 PM
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zerrin responded with a statement of type: Question
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why don't the flame is open? Don't you consider the contamination risk, or do you think that it is not necessary?? |
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| | 10/22/2008 8:11:51 PM
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kamal prajapati responded with a statement of type: Neutral
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That was indeed a great thing for the beginners in this field. Good job! |
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| | 11/10/2008 4:45:38 AM
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Andrew the Clown responded with a statement of type: Neutral
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Did anyone else catch the masive fart at 6:20? It kinda surprised me there! |
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