Cloning-Isolation of Sequencing Grade Plasmid | PraxiLabs

Cloning-Isolation of Sequencing Grade Plasmid Virtual Lab Simulation

Biology | Biochemistry | Genetics | Microbiology



As Featured In

General Aim

To isolate high quality sequencing grade plasmid DNA.

Method

SDS/alkaline lysis.

Learning Objectives (ILO’s)

  • To Apply the steps of high-quality DNA isolation.

  • To understand the role of the devices, the reagents, and the enzymes used in the process of DNA extraction.

  • To implement proper storage of samples.

Theoretical Background/Context

Highly purified plasmid DNA from Escherichia coli are required to obtain reliable sequencing results. This purification is based on the differential denaturation of chromosomal and plasmid DNA. 

Lysis solution containing sodium dodecyl sulfate (SDS) and sodium hydroxide denatures both chromosomal and plasmid DNA in bacterial culture. Neutralization with potassium acetate allows only plasmid DNA to reanneal and stay solubilized. Most of the chromosomal DNA and proteins precipitate in a complex formed with potassium and SDS.

 

Principle of Work

Cells are subjected to SDS/alkaline lysis, to break down the cell wall and extract DNA into the solution. The resulting lysate is neutralized and then centrifuged to pellet out cell debris and the SDS precipitate. The lysate containing the plasmid DNA is applied to the spin column containing the silica membrane, upon which the plasmid DNA will bind. After washing of the silica membrane to remove any contaminants, the plasmid DNA is eluted in a small volume of buffer.


PraxiLabs is Recognized Worldwide

Customers Love PraxiLabs

“With the onset of the COVID-19 pandemic, we found ourselves in a situation that forced us to act quickly to find the best solution available to provide our students with a quality molecular genetics laboratory experience.”

Korri Thorlacius, B.Sc.
Biology Laboratory Instructor
Biology Department
Kwantlen Polytechnic University

'' Although there are now several vendors offering virtual reality software for physics labs, there is only one that offers a realistic, I feel like I’m in a real lab, solution: PraxiLabs.''

Dr.‌ ‌William‌ ‌H.‌ ‌Miner,‌ ‌Jr.‌ ‌
Professor‌ ‌of‌ ‌Physics‌ ‌
Palm‌ ‌Beach‌ ‌State‌ ‌College‌ ‌
Boca‌ ‌Raton,‌ ‌FL‌

" PraxiLabs offered my students a chance to actively engage with the material. Instead of watching videos on a topic, they could virtually complete labs and realize the practical applications of class topics. This is a quality alternative to in-person labs."

Crys Wright
Teaching Assistant
Texas A&M University, USA

"Great user experience and impressive interaction, I am very pleased to have tried the simulations and will continue to do so."

Dr. Khaled M Goher
Lecturer in Biomedical Engineering
Aston University, UK

The #1 Science Virtual Labs used by Educational Institutions

Explore More Interactive 3D Virtual Simulations

Designed for Safety and Engagement

Find out how PraxiLabs keeps students engaged and improves learning outcomes