Gene Cloning Flashcards
What is cloning?
the process of making a genetically identical copy of something
What is gene cloning?
The process of making an exact copy of a particular gene
Why would it be useful to be able to make an exact copy of a particular gene?
usually because of the protein it codes
What are the two main methods of gene cloning?
- In vitro
- In vivio
What does “in vitro” mean?
many copies of a target gene can be made by combining chemicals in a test tube
What is the main technique used to clone genes “in vitro”?
Polymerase chain reaction (PCR)
What is Polymerase Chain Reaction used for in this context?
Polymerase Chain Reaction (PCR) is a technique used to make lots of copies (clones) of a specific region of DNA
What are the three steps of Polymerase Chain Reaction (PCR)?
(D.A.E)
1.Denaturation
2. Annealing
3. Extension
What is the denaturation step of Polymerase Chain Reaction? (with temp conditions)
DNA separates to form single strands at 95 degrees
What is the annealing step of Polymerase Chain Reaction? (with temp conditions)
The primers bind to both DNA strands at 55 degrees
What is the extension step of Polymerase Chain Reaction? (with temp conditions)
The DNA polymerase synthesises new DNA for each strand, suing the orginal strands of DNA like a template at 72 degrees
(so each separated strand is re-fixed with a brand-new, custom match strand so we have two full strands of DNA)
What does “in vivo” mean?
a living organism (like bacteria) is used to make lots of copies of the target gene
What’s the main step of “in vivio” gene cloning?
inserting the target gene into the bacterium (recombinant DNA technology)
What are the basic steps of making recombinant DNA?
- Cutting and pasting the target gene into a plasmid
