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 Korean was automatically generated through Google Translate.
English Version | Other Languages

 JoVE General

열 충격 방법을 사용 E. 대장균에 플라스미드 DNA의 변화

,

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: 50.17.109.248, User IP: 50.17.109.248, User IP Hex: 840003064

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: 열 충격 방법을 사용 E. 대장균에 플라스미드 DNA의 변화

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: 열 충격 방법을 사용 E. 대장균에 플라스미드 DNA의 변화

열 충격 방법을 사용하여 E. 대장균에 플라스미드 DNA의 변화 분자 생물학의 기본 기술입니다. 그것은 박테리아에 외국 플라스미드 또는 결합 제품을 삽입 구성되어 있습니다. 이 비디오 프로토콜은 Genlantis에서 상용 화학적으로 유능한 박테리아를 사용하여 변화의 전통적인 방법을 설명합니다. 얼음에 짧은 부화 후, 화학적 능력 박테리아와 DNA의 혼합물은 45 초 동안 42 ° C에 배치됩니다 (열충격) 후 얼음에 다시 배치. SOC 미디어가 추가되고 변형 세포 ° C 30 분 교반과 함께 37 incubated 수 있습니다. 관계없이 변환 효율의 식민지를 고립을 보장하기 위해서, 변형 박테리아의 두 수량은 도금 있습니다. 이것은 전통적인 프로토콜은 대부분의 상용 유능한 박테리아를 변환하기 위해 성공적으로 사용될 수 있습니다. Genlantis에서 turbocells이 설명서에 설명된 소설 3 분 변환 프로토콜에도 사용할 수 있습니다.

Disclosures: 열 충격 방법을 사용 E. 대장균에 플라스미드 DNA의 변화

Ask the Author: 열 충격 방법을 사용 E. 대장균에 플라스미드 DNA의 변화

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 Is that a new technique that has not reached my university yet, or is it an outdated technique that scientists don't use anymore?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