MILT 525 EXAM 2 FINAL PAPER 2026
QUESTIONS WITH SOLUTIONS GRADED A+
◉ Nuclease. Answer: enzyme that degrades nucleic acids via
hydrolyzing and ester bond
◉ Tm. Answer: Temperature at which 50% of the DNA strand is
denatured
◉ Agarose Gel Electrophoresis. Answer: separates DNA molecules by
size
-EtBr for staining
◉ Blotting steps. Answer: 1. isolate DNA (restriction enzymes)
2. Run electrophoresis
3. Transfer to blot
4. Hybridize with DNA probe
,◉ PCR steps. Answer: 1. Denaturation 30s
2. Annealing 30s
3. Extension 30s then 5 mins
Run product on agarose gel and stain with EtBr
◉ PCR requires (5). Answer: -Template DNA
-Primers
-dNTPs
-Taq polymerase
-bugger (Mg2+)
◉ RT-PCR. Answer: Uses Reverse transcriptase to create cDNA for
PCR.
-measures RNA expression levels
◉ qPCR (Real-time). Answer: Detects PCR products in real time
-Probes that fluoresce is used for detection
-detection during exponential phase of PCR
, -Optical module detects fluorescence and computer quantifies the
data
◉ qPCR probes/dyes. Answer: TaqMan - 5' reporter is cleaved off for
fluorescence
SYBR green Dye - binds to dsDNA and fluoresces
◉ Structure of DNA polymer. Answer:
◉ Mutations in DNA. Answer: Substitution - replacing a base
Insertion - adding a base
Deletion - removing a base
◉ Framshift. Answer: insertion or deletion in the CODING REGION
that alters the reading frame
◉ Null mutation. Answer: results in complete elimination of the
gene (deletion)
QUESTIONS WITH SOLUTIONS GRADED A+
◉ Nuclease. Answer: enzyme that degrades nucleic acids via
hydrolyzing and ester bond
◉ Tm. Answer: Temperature at which 50% of the DNA strand is
denatured
◉ Agarose Gel Electrophoresis. Answer: separates DNA molecules by
size
-EtBr for staining
◉ Blotting steps. Answer: 1. isolate DNA (restriction enzymes)
2. Run electrophoresis
3. Transfer to blot
4. Hybridize with DNA probe
,◉ PCR steps. Answer: 1. Denaturation 30s
2. Annealing 30s
3. Extension 30s then 5 mins
Run product on agarose gel and stain with EtBr
◉ PCR requires (5). Answer: -Template DNA
-Primers
-dNTPs
-Taq polymerase
-bugger (Mg2+)
◉ RT-PCR. Answer: Uses Reverse transcriptase to create cDNA for
PCR.
-measures RNA expression levels
◉ qPCR (Real-time). Answer: Detects PCR products in real time
-Probes that fluoresce is used for detection
-detection during exponential phase of PCR
, -Optical module detects fluorescence and computer quantifies the
data
◉ qPCR probes/dyes. Answer: TaqMan - 5' reporter is cleaved off for
fluorescence
SYBR green Dye - binds to dsDNA and fluoresces
◉ Structure of DNA polymer. Answer:
◉ Mutations in DNA. Answer: Substitution - replacing a base
Insertion - adding a base
Deletion - removing a base
◉ Framshift. Answer: insertion or deletion in the CODING REGION
that alters the reading frame
◉ Null mutation. Answer: results in complete elimination of the
gene (deletion)
