ASCP Molecular Biology Certification Exam
LATEST VERSION WITH COMPLETE
QUESTIONS AND CORRECT DETAILED
ANSWERS \NEWEST EXAM 2026-
VERIFIED 100% ALREADY GRADED A+
What does the incubation step in hybridization do?
Allows formation of ds molecules
Formamide acts as a _____ in a hybridization.
denaturing agent
Line Probe Assay (LiPA)
reverse hybridization assay using sequence-specific oligonucleotide
probes (reverse SSOP)
multi-parameter testing --> single strip
,Single-Stranded Conformational Polymorphism Ananlysis (SSCP)
Used for known gene, unknown mut
Mutation screening
Short PCR products form 3D conformation when cooled --> muts have
different conformation than WT
Non-denaturing PAGE, muts migrate different than WT
How does EtBr cause DNA to fluoresce?
Intercalates into the double
helix Absorbs UV ~300 nm, emits
~600 nm
Pulse Field Gel Electrophoresis steps
1. culture
2. embed pellet in agarose plug
3. treat w/ lysozyme (cell lysis)
4. proteinase K
5. gel
What are the 3 steps of PCR and their temperatures?
Denature 90-
96C Anneal 50-
70C Extension
68-75C
,Bisulfite DNA sequencing/Methylation specific
1. RE digest
2. Electrophorese and purify fragment of interest
3. Denature and incubate w/ sodium bisulfate (turns C>U,
methylated C is unchanged)
4. clean, ppt, and resuspend
5. PCR --> sequence
6. Compare treated vs untreated, note where CG are not changed to TA
NASBA steps
1. Hybridize oligo-T7P primer to target seq
2. RT/RNase H
3. Hybridize with target-specific oligo primer (P2)
4. RNA transcript of T7 RNA pol
Do you have to know the gene sequence in order to do DNA sequencing?
Yes, in order to design primers
You do NOT need to know the mutation
Sanger sequencing method
Divided into 4 samples (ddA, T, G, C)
Label with radioactive/dye oligo at
3' end Mix with taq, dNTPs, ddNTP
and incubate
run on gel --> frags will terminate at different lengths
, Fluorescent in situ hybridization (FISH)
Uses fluorescent probes to detect DNA sequences on chr
What types of probes are used for FISH?
Dual fusion: 2 probes flank the breakpoint at both t locations
CEN probes: centromeric probes bind to repetitive alpha satellite
sequences Telomeric probes
Whole chr paints
What is the wavelength for background in spectrophotometery?
320 nm
How do you determine quality of DNA/RNA using a spectrophotometer?
A260/A280
What is considered good quality DNA/RNA from spectrophotometry?
DNA: 1.7-2.0
RNA: 2.0-2.3
What is considered poor quality RNA/DNA from spectrophotometry?
<1.7 Protein contamination
LATEST VERSION WITH COMPLETE
QUESTIONS AND CORRECT DETAILED
ANSWERS \NEWEST EXAM 2026-
VERIFIED 100% ALREADY GRADED A+
What does the incubation step in hybridization do?
Allows formation of ds molecules
Formamide acts as a _____ in a hybridization.
denaturing agent
Line Probe Assay (LiPA)
reverse hybridization assay using sequence-specific oligonucleotide
probes (reverse SSOP)
multi-parameter testing --> single strip
,Single-Stranded Conformational Polymorphism Ananlysis (SSCP)
Used for known gene, unknown mut
Mutation screening
Short PCR products form 3D conformation when cooled --> muts have
different conformation than WT
Non-denaturing PAGE, muts migrate different than WT
How does EtBr cause DNA to fluoresce?
Intercalates into the double
helix Absorbs UV ~300 nm, emits
~600 nm
Pulse Field Gel Electrophoresis steps
1. culture
2. embed pellet in agarose plug
3. treat w/ lysozyme (cell lysis)
4. proteinase K
5. gel
What are the 3 steps of PCR and their temperatures?
Denature 90-
96C Anneal 50-
70C Extension
68-75C
,Bisulfite DNA sequencing/Methylation specific
1. RE digest
2. Electrophorese and purify fragment of interest
3. Denature and incubate w/ sodium bisulfate (turns C>U,
methylated C is unchanged)
4. clean, ppt, and resuspend
5. PCR --> sequence
6. Compare treated vs untreated, note where CG are not changed to TA
NASBA steps
1. Hybridize oligo-T7P primer to target seq
2. RT/RNase H
3. Hybridize with target-specific oligo primer (P2)
4. RNA transcript of T7 RNA pol
Do you have to know the gene sequence in order to do DNA sequencing?
Yes, in order to design primers
You do NOT need to know the mutation
Sanger sequencing method
Divided into 4 samples (ddA, T, G, C)
Label with radioactive/dye oligo at
3' end Mix with taq, dNTPs, ddNTP
and incubate
run on gel --> frags will terminate at different lengths
, Fluorescent in situ hybridization (FISH)
Uses fluorescent probes to detect DNA sequences on chr
What types of probes are used for FISH?
Dual fusion: 2 probes flank the breakpoint at both t locations
CEN probes: centromeric probes bind to repetitive alpha satellite
sequences Telomeric probes
Whole chr paints
What is the wavelength for background in spectrophotometery?
320 nm
How do you determine quality of DNA/RNA using a spectrophotometer?
A260/A280
What is considered good quality DNA/RNA from spectrophotometry?
DNA: 1.7-2.0
RNA: 2.0-2.3
What is considered poor quality RNA/DNA from spectrophotometry?
<1.7 Protein contamination
