Lecture 6: DNA Techniques

Question 1

Kary Mullis

Answer 1

Developed PCR and revolutionized research because of its ability to create billions of copies (clones) of a given region f DNA only in hours - as opposed to cloning in bacteria

Question 2

Taq DNA Polymerase

Answer 2

• If a standard DNA polymerase was used for PCR, the polymerase would be denatured along with the DNA during the repeated heating steps at 94C
• The key to easy PCR automation was the discovery of the unusual heat-stable DNA Taq Polymerase isolated from a bacterium living in hot springs

Question 2

Taq DNA Polymerase

Answer 2

• If a standard DNA polymerase was used for PCR, the polymerase would be denatured along with the DNA during the repeated heating steps at 94C
• The key to easy PCR automation was the discovery of the unusual heat-stable DNA Taq Polymerase isolated from a bacterium living in hot springs

Question 3

Uses of PCR in Modern Biotechnology

Answer 3

• amplify from one organism a gene that may be similar in sequence to one from another organism
• basic research
• completing sequences in genome projects
• forensics - amplify crime scene DNA
• paternity testing
• rapid diagnosis of genetic disorders
• rapid diagnosis of infectious disease
• protection of habitats and biodiversity

Question 4

Colony Bot

Answer 4

Finding Gene of interest from 1000s of clones from DNA library

Question 5

Southern Blotting- Applications

Answer 5

Southern blotting allows for the detection of specific DNA fragments in a complex DNA mixture
1. confirm the presence of a recombinant DNA in a host organism
2. Diagnosis of mutant alleles in the disease
3. Determine the number of copies of a recombinant DNA gene in a host organism
4. Determine the size of the fragment that contains the sequence
5. Determine the presence of a similar gene in another organism

Question 6

Sequencing Synthesis

Answer 6

Detects fluorescently labeled nucleotides upon incorporation

Question 7

Ion Proton Sequencing

Answer 7

Detects H+

Question 8

Pyrosequencing

Answer 8

detects phosphate