Basic Techniques

Question 1

Essential components for vectors

Answer 1

Origin of replication

Dominant selectable marker (antibiotic resistance gene)

Unique RE sites for gene insertion

Question 2

Southern Blot

Answer 2

Determines presence of specific DNA sequence in a sample using a DNA probe

Question 3

Northern Blot

Answer 3

Determines presence, size, and abundance of RNA using a DNA probe

Used in gene expression studies

Question 4

SDS-Page

Answer 4

(Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis)

Used to separate proteins based on size

Denatures proteins and provides net negative charge

Stain with Coomassie blue to detect abundance of protein (darker= more protein)

Question 5

Immunofluorescence

Answer 5

Uses Ab to detect portein location in fixed cells

Question 6

Immunoprecipitation

Answer 6

Uses AB to isolate a protein for western blot analysis

Question 7

Reverse Transcriptase (RT)

Answer 7

Converts mRNA to cDNA

Question 8

Forward primer

Answer 8

○ Anneals to the bottom strand (has same sequence as the top strand)
Primer extension synthesizes the top strand

Question 9

Reverse primer

Answer 9

○ Anneals to the top strand (has same sequence as the bottom strand)
Primer extension synthesizes the bottom strand

Question 10

Ras Gene Mutation

Answer 10

§ Glycine 12 on WT gene changed to valine

Presence of valine turns cancerous gene “on”