GENETICS EXAM 4 COMPLETE COURSE
REVIEW GUIDE
◉ Restriction enzymes.
Answer: Cut DNA at specific recognition sites; used to generate
compatible ends for cloning and to analyze DNA fragments.
◉ Gel electrophoresis.
Answer: Separates DNA fragments by size in an agarose gel under an
electric field.
Smaller fragments migrate farther.
◉ PCR.
Answer: amplification of a specific DNA region via:
1. Denaturation (template strands separate)
2. Annealing (primers bind)
3. Extension (Taq polymerase synthesizes new strands)
Produces exponential amplification. Requires template, primers,
dNTPs, Taq, buffer.
◉ Transformation of a bacterial cell.
Answer: introduction of foreign DNA into bacterial cells
, ◉ important components of a cloning vector.
Answer: A plasmid containing an origin of replication, selectable
marker, and MCS that allows propagation of inserted DNA.
◉ How does screening for successful transformation and inserted
target DNA work.
Answer: Method using disruption to identify bacteria with
successful DNA insertions (white colonies have inserts
◉ Expression Vector Prokaryotic.
Answer: bacterial promoter, Shine-Dalgarno sequence, prokaryotic
transcription terminators.
◉ Expression Vector Eukaryotic.
Answer: eukaryotic promoter, Kozak sequence for translation
initiation, polyA signal, and introns sometimes required for stable
expression.
◉ DNA vs. cDNA.
Answer: DNA contains introns, promoters, and noncoding regions;
cDNA is reverse-transcribed from mature mRNA and contains only
expressed exons.
REVIEW GUIDE
◉ Restriction enzymes.
Answer: Cut DNA at specific recognition sites; used to generate
compatible ends for cloning and to analyze DNA fragments.
◉ Gel electrophoresis.
Answer: Separates DNA fragments by size in an agarose gel under an
electric field.
Smaller fragments migrate farther.
◉ PCR.
Answer: amplification of a specific DNA region via:
1. Denaturation (template strands separate)
2. Annealing (primers bind)
3. Extension (Taq polymerase synthesizes new strands)
Produces exponential amplification. Requires template, primers,
dNTPs, Taq, buffer.
◉ Transformation of a bacterial cell.
Answer: introduction of foreign DNA into bacterial cells
, ◉ important components of a cloning vector.
Answer: A plasmid containing an origin of replication, selectable
marker, and MCS that allows propagation of inserted DNA.
◉ How does screening for successful transformation and inserted
target DNA work.
Answer: Method using disruption to identify bacteria with
successful DNA insertions (white colonies have inserts
◉ Expression Vector Prokaryotic.
Answer: bacterial promoter, Shine-Dalgarno sequence, prokaryotic
transcription terminators.
◉ Expression Vector Eukaryotic.
Answer: eukaryotic promoter, Kozak sequence for translation
initiation, polyA signal, and introns sometimes required for stable
expression.
◉ DNA vs. cDNA.
Answer: DNA contains introns, promoters, and noncoding regions;
cDNA is reverse-transcribed from mature mRNA and contains only
expressed exons.
