Intro to Genetic Assays & Cell Cycle

Question 1

Polymerase Chain Reaction

Answer 1

Used to amplify DNA; Repeated copying of a target sequence of DNA; Taq polymerase; Primers (required to initiate elongation) target the sequence that needs to be copied; Thermal cycling; Only needs trace amounts of DNA

Question 2

Restriction Endonucleases

Answer 2

Binds to specific sequences of DNA; Short, palindromic sequences; Can produce “sticky” or “blunt” ends; Type II is most commonly used because of predictable cutting; over 200 endonucleases, each recognizing a specific DNA sequence

Question 3

Restriction Fragment Length Polymorphism

Answer 3

Differences in DNA sequences can generate different DNA cleavage sites for restriction enzymes; Two different alleles will produce different fragment patterns

Question 4

Electrophoresis

Answer 4

Separates DNA and RNA fragments based on size; Smaller fragments migrate farther than larger, heavier fragments

Question 5

Types of directional blots

Answer 5

Southern: DNA
Nothern: RNA
Western: Protein
Eastern: Posttranslational modifications

Question 6

Southern blot

Answer 6

Extract DNA and digest it with restriction enzymes; Run electrophoresis; Bands on gel are blotted onto a membrane; Radioactive probe that hybridizes with the complementary DNA sequence; Expose to x-ray films; Probe labels different restriction enzymes

Question 7

Northern blot

Answer 7

Extract RNA; Electrophoresis; Transfer RNA fragments to membrane and fix with UV or heat; Membrane hybridized with labeled probes; Thicker the band, higher level of expression

Question 8

DNA Microarray

Answer 8

Analyzes active transcription; Uses cDNA; Simultaneous analysis of various genes

Question 9

Chain Termination DNA Sequencing (Sanger Method)

Answer 9

Initiate strand synthesis; Add dideoxynucleotide which stops strand synthesis; Run electrophoresis

Question 10

Immunoassays

Answer 10

Biochemical test that measures the presence or concentration of a macromolecule in a solution

Question 11

Cell Cycle Check-Pts

Answer 11

G1, G2, Spindle Assembly