Gel Electrophoresis Flashcards
outline the safety considerations when setting up and running a gel electrophoresis analysis
personal protective equipment:
- lab coat, gloves, safety goggles
handling of chemicals, limiting electrical hazards, handling DNA samples, correct waste disposal, general lab safety practices.
explain how the enzymes endonuclease and ligase can be used to create a recombinant plasmid
endonuclease are restriction enzymes, and they play a vital role in the creation of a recombinant plasmid:
- they are used to cut the plasmid vector and insert DNA at specific restriction sites.
- these cuts create compatible sticky and blunt ends which allows for the plasmid and insert to be joined together.
DNA ligase plays a crucial role in the final step of creating a recombinant plasmid:
- DNA ligase is used to seal the plasmid and insert DNA after they have been cut by restriction enzymes.
- the ligase forms phosphodiester bonds between the compatible ends of the plasmid and insert DNA, ensuring the two DNA pieces are covalently joined together.
- the result is a recombinant plasmid
discuss the advantages of using bacterial plasmids to produce human proteins
using bacterial plasmids in order to produce human proteins prove advantageous in many aspects:
- simplicity of genetic manipulation»_space; bacteria is easy to manipulate.
- cost-effectiveness»_space; production is at high rates
- low risk of contamination»_space; using human pathogens decreases the risk.
Outline the steps required to place a recombinant plasmid back into bacteria via heat shock
- Prepare bacteria cells
- Mix plasmid DNA
- Heat shock the mixture
- Return to ice
- Plate cells on agar plates
- Screen colonies
