The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

Recommend to Librarian

Automatic Translation

This translation into Swedish was automatically generated through Google Translate.
English Version | Other Languages

 JoVE General

Omvandling av Plasmid DNA i E. coli med metoden Heat Shock

,

Department of Physiology and Biophysics, University of California, Irvine (UCI)

You must be subscribed to JoVE to access this content.

This article is a part of   JoVE General. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.

Recommend JoVE to Your Librarian

Current Access Through Your IP Address

You do not have access to any JoVE content through your current IP address.

IP: 50.16.108.167, User IP: 50.16.108.167, User IP Hex: 839937191

Current Access Through Your Registered Email Address

You aren't signed into JoVE. If your institution subscribes to JoVE, please or create an account with your institutional email address to access this content.

 

Video Article Chapters

Cite this Article: Omvandling av Plasmid DNA i E. coli med metoden Heat Shock

Froger, A., Hall, J. E. Transformation of Plasmid DNA into E. coli Using the Heat Shock Method. J. Vis. Exp. (6), e253, doi:10.3791/253 (2007).

Abstract: Omvandling av Plasmid DNA i E. coli med metoden Heat Shock

Omvandling av plasmid DNA i E. coli med metoden värmechock är en grundläggande teknik för molekylärbiologi. Den består av föra in en främmande plasmid eller ligatur produkt i bakterier. Denna video protokoll beskriver den traditionella metoden för omvandling med hjälp av kommersiellt tillgängliga kemiskt behöriga bakterier från Genlantis. Efter en kort inkubation i is, är en blandning av kemiskt behöriga bakterier och DNA placeras på 42 ° C i 45 sekunder (heat shock) och placeras sedan tillbaka i is. SOC media läggs och förvandlas cellerna inkuberas vid 37 ° C i 30 min med agitation. För att vara säker på att isolera kolonier oavsett omvandlingseffektiviteten är två mängder förvandlade bakterier pläterade. Denna traditionella protokollet kan användas framgångsrikt för att omvandla de flesta kommersiellt tillgängliga behöriga bakterier. Den turbocells från Genlantis kan också användas på ett nytt 3-minuters omvandling protokoll, som beskrivs i bruksanvisningen.

Disclosures: Omvandling av Plasmid DNA i E. coli med metoden Heat Shock

Ask the Author: Omvandling av Plasmid DNA i E. coli med metoden Heat Shock

9 Comments

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?

1

Reply

Posted by: AnonymousFebruary 1, 2008, 7:57 PM

hi my neam isalirezababazade . iam student in unversity tehran . what do you teransfer factors F(negativ)in the e.coli?why transfe geneboth factor?

1

Reply

Posted by: aliDecember 24, 2008, 9:41 AM

People use it all the time. Ask in the labs around, I am sure you will find people who use it.

1.1

Reply

Posted by: AnonymousFebruary 1, 2008, 8:11 PM

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? 

1.2

Reply

Posted by: AnonymousMarch 9, 2008, 8:45 PM

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)

1.3

Reply

Posted by: ozgur g.September 9, 2008, 9:04 PM

chemical dna transformation and plasmid curing of E.coli

1.4

Reply

Posted by: ali sabahJanuary 11, 2012, 11:49 PM

why don't the flame is open? Don't you consider the contamination risk, or do you think that it is not necessary??

3

Reply

Posted by: zerrinSeptember 9, 2008, 4:22 PM

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.   

3.1

Reply

Posted by: labSeptember 16, 2008, 8:57 PM

That was indeed a great thing for the beginners in this field. Good job!

6

Reply

Posted by: kamal prajapatiOctober 22, 2008, 8:11 PM

double stranded plasmid can be fntered into host

9

Reply

Posted by: khemJune 10, 2010, 6:00 AM

Beads are used as well as scoops, i prefare scoops because i can be sure that everything is spread nice and equal. some technician use beads, you need to do the right motion to make it work :D

10

Reply

Posted by: MuadJanuary 25, 2012, 3:59 AM

Why we are keep the mixture of competent cells and plasmid DNA at 42c for 45 minutes?

11

Reply

Posted by: NagarjunaFebruary 22, 2012, 8:56 AM

45 seconds not minutes XD
Useful, more protocols should be explain like this one
thaks

12

Reply

Posted by: AdrianApril 12, 2012, 7:48 PM

Where is gloves to prevent contamination?????

13

Reply

Posted by: OHIONET O.May 10, 2012, 4:35 PM

Post a Question / Comment / Request

You must be signed in to post a comment. Please or create an account.

Waiting
simple hit counter