Fundamentals of Biology

Agarose Gel Electrophoresis, DNA Sequencing, PCR

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Session Overview

This session will outline methods to analyze genes identified through recombinant DNA technologies. Learning Objectives To understand what an agarose gel is and how to use agarose gel electrophoresis to analyze DNA molecules. To understand the basic mechanism of DNA sequencing by the dideoxy chain termination method. To know that there is a vast database containing the DNA sequence of the entire genomes for many different organism, and understand why this is useful. To understand the basic mechanism of the polymerase chain reaction (PCR) method of amplifying DNA and know when to use this technique.

Session Activities

Lecture Video

Watch the lecture video excerpt

• Agarose Gel Electrophoresis, DNA Sequencing, PCR, Excerpt 1

Check Yourself

Question 1

Select the true statement or statements.

Agarose gel electrophoresis separates DNA molecules by size. check

When using Agarose gel electrophoresis, DNA molecules move toward the negative electrode. close

When using Agarose gel electrophoresis, large DNA molecules move faster through the gel. close

Check

Question 2

When using chain termination sequencing, the nucleotides added as a chain terminator are…

missing the hydroxyl on both the 2’ and 3’carbon check

missing the hydroxyl on the 2’ carbon close

missing the hydroxyl on the 3’ carbon close

Check

Session Activities

Lecture Video

Watch the lecture video excerpt

• Agarose Gel Electrophoresis, DNA Sequencing, PCR, Excerpt 2

Check Yourself

Question 1

Which of the following are required for a successful PCR amplification? 1. One or more copies of the DNA to be amplified. 2. all of the normal DNA nucleotides. 3. some dideoxynucleotides 4. Two distinct primers 5. E. coli DNA polymerase 6. DNA polymerase from an organism that lives at high temperature

All of these are required close

a, b, d, f check

a, c, d, e close

a, c, d, f close

b, c, d, e close

Check

Session Activities

Help Session Video

Watch the short video of Robert Dorkin explaining the Polymerase Chain Reaction (PCR) and DNA sequencing.

• Polymerase Chain Reaction (PCR)

Practice Problems

• Problems (PDF)
• Solutions (PDF)

Further Study

Useful Links

• DNA Sequencing
• Jonathan Weissman: DNA Sequencing
• Biology Animation Library
• PCR Virtual Lab

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