Unit 4 - Ch 15 - Biotechnology

Question 1

DNA Sequencing

Answer 1

Determine the nucleotide sequence in DNA

• highly automated
• human genome project
• databases online
• computational biology / bioinformatics

Question 2

Restriction Endonucleases

Answer 2

Enzymes isolated from bacteria used by bacteria to “fight” off bacteriophages (“bacteria eaters”)

• cleave DNA at specific sequences

Question 3

Gel Electrophoresis

Answer 3

Uses a gel matrix to separate fragments of DNA

• separates based upon size of fragments
• apply electrical current

Question 4

RFLP steps and uses

Answer 4

Restriction-Fragment Length Polymorphism

1. Steps
   a. Fragment DNA - via restriction enzyme
   b. Separate fragments - via electrophoresis
   c. Compare w/ known samples
2. Uses
   a. Paternity Suits
   b. Criminal Investigations

Question 5

PCR

Answer 5

Polymerase Chain Reaction

• used to amplify DNA (copy)
• uses thermal cycler (3 temp phases)
• Taq polymerase from thermus aquaticus from Yellowstone

Question 6

CRISPR

Answer 6

Clusters of Regularly Interspaced Short Palindromic Repeats

• sequences from viral DNA found in bacteria
• Cas9- RNA guided endonuclease - guide RNA, detect sequence of interest (cleavage)