Molecular Biology

Question 1

DNA/RNA analysis?

Answer 1

DNA/RNA isolation – disrupts cell membranes – gel electrophoresis (method
for separation + analysis of macromolecules (DNA, RNA + proteins) + their
fragments, based on their size + charge.

• PCR- polymerase chain reaction: (used to amplify single copy or few copies of
  a piece of DNA, generating thousands to millions of copies of a particular DNA
  sequence).
  Denaturation of DNA- heating reaction to 95°C to break H-bonds between double
  stranded DNA.
  Primer annealing- temp lowered to 60°C to allow primers (w. complimentary base
  pairing) anneal to end of strand using polymerase.
  Elongation- temp increased to 72°C so that DNA polymerase can synthesize new
  DNA strand complementary to DNA template strand by adding nucleotides that
  are complementary to template in 5’ to 3’ direction

Question 2

Restriction analysis?

Answer 2

restriction endonucleases (uses enzymes to cleave DNA
at specific sites along the molecule

Question 3

DNA sequencing?

Answer 3

determining the precise order of nucleotides within DNA

molecule. Peak for each base on chromatogram

Question 4

Hybridization?

Answer 4

detection of specific sequence w. labelled probe.
probe is single stranded polynucleotide w. known sequence + is labelled w.
radioactive elements, fluorescent dyes.
Probe hybridizes to complimentary DNA
sequence and glows or visualization (radioactivity, fluorescence or
chemiluminiscence)

Question 5

Southern blot?

Answer 5

hybridization of genomic DNA

Question 6

Northern blot?

Answer 6

mRNA

hybridization

Question 7

DNA fingerprinting?

Answer 7

analyses of non-coding regions of genome

Question 8

Medical Applications?

Answer 8

detection of inherited mutations/genetic disorders
detection of acquired mutations/oncogenetics
detection of pathogens causing infections/diseases
detection of mutations

Question 9

Therapy?

Answer 9

recombinant proteins