Integrative Biology 100
Discussion Class, March  31- April 2
#10 Recombinant DNA
and Polymerase Chain Reaction (PCR)

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Related Assignments:

Third Moodle Assignment due by 8:00 AM Thursday April 8 

Take-home Assignment #7 Control of Gene Expression & Biotechnology due at Lecture Tuesday April 13 

Learner Objectives

You should be able to:

1. Draw a diagram or write a description of the action of restriction enzymes in the cutting of DNA molecules into "restriction fragments".

2. Outline a procedure for isolating a specific gene from a eukaryotic organism, incorporating it into a recombinant plasmid, and inserting it into a bacterial cell to produce a gene product.

donor DNA	restriction enzyme
restriction site	sticky ends
plasmid	recombinant DNA
ligase	vector
Promotor

3. Describe how the polymerase chain reaction (PCR) is used to make millions of copies of specific sequences of DNA. Discuss the significance of PCR in the location of specific DNA sequences in a small sample of tissue.

PCR polymerase chain Reaction	Thermus aquaticus and taq polymerase
Yellowstone National Park	high temperatures
unzipping DNA	DNA primers for specific DNA sequences
DNA nucleotides	binding or annealing of primers
DNA replication	repetition

Web Resources for this discussion:

DNA Interactive

Recombinant DNA

Select Manipulation from the left-hand menu

Choose "Techniques" at the bottom of the next screen

Choose "Cutting and pasting" at the top of the next screen

Two animations, "Cutting and pasting DNA" and "Recombining DNA" are very useful.

Polymerase Chain Reaction

• Animation of the Polymerase Chain Reaction from sumanasinc.com.
  Select "The Polymerase Chain Reaction (PCR) ".
  Just watch the animation to get the "gist" of the concept of PCR.

• *Polymerase Chain Reaction from DNA Interactive

Choose "Techniques" at the bottom of the next screen

Choose "Amplifying" at the top of the next screen

Two animations, "Making many copies of DNA" and "PCR animation" are very useful.