MB (ASCP) EXAM QUESTIONS AND ANSWERS
Consider the table below where a child and a possible father (PF) share the listed
paternity indices for each locus listed (LOC-A1, LOC-B2, LOC-C3, LOC-D4).
Locus Tested PF Child Paternity Index
LOC-A1 3 2/3 2.18
LOC-B2 7/5 5 0.798
LOC-C3 15/17 9/17 5.21
LOC-D4 12 12 1.37
Based on the data presented in the table, what is the combined paternity index, CPI,
from the loci tested: LOC-A1, LOC-B2, LOC-C3 and LOC-D4?
12.42
9.558
15.56
2.38 - Answers -12.42 ( CPI is calculated by multiplying all Paternity index tgt)
Best way to avoid PCR contamination - Answers -Separate Pre-PCR (clean area) and
post PCR (dirty area)
A260=1.4, A280=1.1 . protein contamination or Rna contam or useable RNA - Answers
-Protein contamination . A260/280 = 1.3
<1.6 = protein contamination, 1.7-2.0 = good DNA, 1.9-2.0 = good RNA
Indexing is performed in NGS - Answers -To Barcode/Label/Tag DNA Sample of
Interest
Ideal Workflow Next Generation Sequencing (NGS) - Answers -Fragmentation-End
Repair-Adapter Ligation
What prevents/quenches the Fluorescent (Report Dye) Signal in a Taq Man Probe -
Answers -G On 3' End
A List of PCR Components for PCR Reaction Mix Provided (Determine which
component is excess): - Answers -DNA Polymerase Concentration needs to be reduced
to 2.5-3 units - Taq Polymerase Conc was provided as 100 Units as compared to other
PCR Components
How to Preferentially amplify short DNA Fragments - Answers -Increase KCL
Concentration
Degenerate Primer Sets - Answers -MultiPlex PCR
,What PCR increases Specificity and sensitivity of a low copy number target in a patient
sample - Answers -Nested PCR
Pressure in pre-area and post-area PCR - Answers -Pre - Positive pressure
Post - Negative pressure
Bake Glassware to be used for RNA Isolation Procedure - Answers -250 C For 4 Hours
(However Book States 4-6 Hours at 400 C)
Isochromosomes - Answers -Long Arms
Term for abnormal number of chromosomes in a cell. For example a human cell having
45 or 47 chromosomes instead of the usual 46. - Answers -Aneuploidy
( A cell with any number of complete chromosome sets is called a euploid cell.)
Nucleic Acid Dyes that binds to minor groove of Double Helix DNA - Answers -SYBR
Green
Addition of Agarose in Gel - Answers -Slows the migration by 15%
Polyacrylamide - Answers -Neurotoxin (Acrylamide Neurotoxin in Gel)
Codon (Nucleotide) + Amino Acid Code + Name of the Amino Acid Provided + Identify
the Kind of Mutation - Answers -Frame Shift Mutation (Shifts the Frame)
Non-Sense Mutation
Interaction between DNA/Histones - Answers -Hydrophobic Interactions
Type of RNA that controls Gene Expression, protein synthesis, Cell Development and
Eukaryotic Cell Defense - Answers -miRNA
mRNA Stability & Half-life - Answers -3' Poly A Tail (5' Methylated G Cap was not an
option)
Type of RNA that serves as template for Transcription Process - Answers -Template
(Anti sense Strand)
Eukaryotic DNA Polymerase for Leading Strand - Answers -δ (Delta)
Eukaryotic DNA Polymerase for Repair - Answers -ε (Epsilon) - (Since Beta {β}
Polymerase was not an option)
How many SNPS are there? - Answers -10^7 or 11 bilion
How to create synthetic dsDNA? - Answers -SDA
,Culture of fibroblasts? - Answers -Trypsin and scrape
Monitoring HIV circulating in patient's plasma using - Answers -NASBA
HIV genome is RNA, so only NABSA would allow direct detection
What is a metagenome? - Answers -sum total of all DNA sequenced from an
environmental sample
What gel to use for 200-300 base pairs? - Answers -0.5 agarose or 8% polyacrylamide
(Pulse Field >25000bp
PAGE 5-500 bp
Agarose 200-30000bp)
What does G574* mean? - Answers -Termination after the G
How to store RNA for more than 6 months? - Answers -Ethanol -70C
How to check for HER2? - Answers -FISH
What causes smearing in gel (high/low dye concentration, high low voltage) - Answers -
High voltage
Improperly prepared gel: If the gel is not poured correctly, it will not polymerize or
solidify evenly, thus causing the molecules to smear.
If the wells are filled too much, or if the sample is not properly diluted, the excess
sample may smear across the gel. Also DNA degradation.
What is associated with multiple mutations and large deletions - Answers -DMD
(Duchenne Muscular Dystrophy) _Dystrophin gene
IQCP is most associated with - Answers -risk assessment
Is Linkage Disequilibrium to nonrandom association of FAR or NEAR loci? - Answers -
Near. (Linkage Disequilibrium occurs when 2 alleles are Close in the same
chromosome. Linkage Equilibrium occurs when 2 alleles are Far apart in the same
chromosome)
Example of ISOTHERMAL SIGNAL amplification?
Technique employs the signal amplification resulting from probe: target hybridization
rather than by amplifying the target or the probe? - Answers -bDNA / bDNA analysis
want to PCR HLA-D - Answers -Beta Exon 2
HCV most conserved - Answers -5'UTR
Mantle (MCL) - Answers -t (11;14)
, Histones - Answers -Ionic
HER2 overexpression - Answers -Treatment with Traustmaub
What is FLT3 associated with? - Answers -AML
HFE - Answers -C282Y, H63D, S65C
Acute Promylocytic Leukemia (APL) - Answers -t(15;17)
Something about getting quality of RNA? Choices were all 3 letter acronym with R, like
RFN, RIN - Answers -The RNA integrity number (RIN) is an algorithm for assigning
integrity values to RNA measurements. The integrity of RNA is a major concern for
gene expression studies and traditionally has been evaluated using the 28S to 18S
rRNA ratio, a method that has been shown to be inconsistent.
Amplification method with polymerase creating a synthetic dsDNA - Answers -TBC
Identical twins for organ donor what should you do? - Answers -continue with assay, tell
supervisor, something about parents?
Why does NGS use fluorescence over spectrometry? - Answers -Fluorescence is more
sensitive because of the different ways of measuring absorbance and fluorescence.
Light absorbance is measured as the difference in intensity between light passing
through the reference and the sample. In fluorescence the intensity is measured
directly, without comparison with a reference beam.
Similarities between NGS and CE/Sanger - Answers -•DNA polymerase incorporates
fluorescently labeled dNTPs into a DNA template
•The newly incorporated nucleotides are identified by fluorophore excitation.
MALDI-Tof - Answers -Matrix-Assisted Laser Desorption/Ionization-Time Of Flight
(MALDI-TOF) Mass Spectrometry (MS) is a common method used for quality control
(QC) of oligonucleotides. MALDI-TOF instrumentation is suitable for use in high-
throughput industrial QC settings and can resolve molecules in the size range of
oligonucleotides
Heteroduplex - Answers -are dsDNA complexes - consisting of two partly mismatched
polynucleotide strands derived from two different parent molecules. (look up Denaturing
HPLC)
Benefits of NGS Output - Answers -Reads are relatively short, but so much more is
produced (100K-1 Billion) creating a huge amount of data for very little cost
Consider the table below where a child and a possible father (PF) share the listed
paternity indices for each locus listed (LOC-A1, LOC-B2, LOC-C3, LOC-D4).
Locus Tested PF Child Paternity Index
LOC-A1 3 2/3 2.18
LOC-B2 7/5 5 0.798
LOC-C3 15/17 9/17 5.21
LOC-D4 12 12 1.37
Based on the data presented in the table, what is the combined paternity index, CPI,
from the loci tested: LOC-A1, LOC-B2, LOC-C3 and LOC-D4?
12.42
9.558
15.56
2.38 - Answers -12.42 ( CPI is calculated by multiplying all Paternity index tgt)
Best way to avoid PCR contamination - Answers -Separate Pre-PCR (clean area) and
post PCR (dirty area)
A260=1.4, A280=1.1 . protein contamination or Rna contam or useable RNA - Answers
-Protein contamination . A260/280 = 1.3
<1.6 = protein contamination, 1.7-2.0 = good DNA, 1.9-2.0 = good RNA
Indexing is performed in NGS - Answers -To Barcode/Label/Tag DNA Sample of
Interest
Ideal Workflow Next Generation Sequencing (NGS) - Answers -Fragmentation-End
Repair-Adapter Ligation
What prevents/quenches the Fluorescent (Report Dye) Signal in a Taq Man Probe -
Answers -G On 3' End
A List of PCR Components for PCR Reaction Mix Provided (Determine which
component is excess): - Answers -DNA Polymerase Concentration needs to be reduced
to 2.5-3 units - Taq Polymerase Conc was provided as 100 Units as compared to other
PCR Components
How to Preferentially amplify short DNA Fragments - Answers -Increase KCL
Concentration
Degenerate Primer Sets - Answers -MultiPlex PCR
,What PCR increases Specificity and sensitivity of a low copy number target in a patient
sample - Answers -Nested PCR
Pressure in pre-area and post-area PCR - Answers -Pre - Positive pressure
Post - Negative pressure
Bake Glassware to be used for RNA Isolation Procedure - Answers -250 C For 4 Hours
(However Book States 4-6 Hours at 400 C)
Isochromosomes - Answers -Long Arms
Term for abnormal number of chromosomes in a cell. For example a human cell having
45 or 47 chromosomes instead of the usual 46. - Answers -Aneuploidy
( A cell with any number of complete chromosome sets is called a euploid cell.)
Nucleic Acid Dyes that binds to minor groove of Double Helix DNA - Answers -SYBR
Green
Addition of Agarose in Gel - Answers -Slows the migration by 15%
Polyacrylamide - Answers -Neurotoxin (Acrylamide Neurotoxin in Gel)
Codon (Nucleotide) + Amino Acid Code + Name of the Amino Acid Provided + Identify
the Kind of Mutation - Answers -Frame Shift Mutation (Shifts the Frame)
Non-Sense Mutation
Interaction between DNA/Histones - Answers -Hydrophobic Interactions
Type of RNA that controls Gene Expression, protein synthesis, Cell Development and
Eukaryotic Cell Defense - Answers -miRNA
mRNA Stability & Half-life - Answers -3' Poly A Tail (5' Methylated G Cap was not an
option)
Type of RNA that serves as template for Transcription Process - Answers -Template
(Anti sense Strand)
Eukaryotic DNA Polymerase for Leading Strand - Answers -δ (Delta)
Eukaryotic DNA Polymerase for Repair - Answers -ε (Epsilon) - (Since Beta {β}
Polymerase was not an option)
How many SNPS are there? - Answers -10^7 or 11 bilion
How to create synthetic dsDNA? - Answers -SDA
,Culture of fibroblasts? - Answers -Trypsin and scrape
Monitoring HIV circulating in patient's plasma using - Answers -NASBA
HIV genome is RNA, so only NABSA would allow direct detection
What is a metagenome? - Answers -sum total of all DNA sequenced from an
environmental sample
What gel to use for 200-300 base pairs? - Answers -0.5 agarose or 8% polyacrylamide
(Pulse Field >25000bp
PAGE 5-500 bp
Agarose 200-30000bp)
What does G574* mean? - Answers -Termination after the G
How to store RNA for more than 6 months? - Answers -Ethanol -70C
How to check for HER2? - Answers -FISH
What causes smearing in gel (high/low dye concentration, high low voltage) - Answers -
High voltage
Improperly prepared gel: If the gel is not poured correctly, it will not polymerize or
solidify evenly, thus causing the molecules to smear.
If the wells are filled too much, or if the sample is not properly diluted, the excess
sample may smear across the gel. Also DNA degradation.
What is associated with multiple mutations and large deletions - Answers -DMD
(Duchenne Muscular Dystrophy) _Dystrophin gene
IQCP is most associated with - Answers -risk assessment
Is Linkage Disequilibrium to nonrandom association of FAR or NEAR loci? - Answers -
Near. (Linkage Disequilibrium occurs when 2 alleles are Close in the same
chromosome. Linkage Equilibrium occurs when 2 alleles are Far apart in the same
chromosome)
Example of ISOTHERMAL SIGNAL amplification?
Technique employs the signal amplification resulting from probe: target hybridization
rather than by amplifying the target or the probe? - Answers -bDNA / bDNA analysis
want to PCR HLA-D - Answers -Beta Exon 2
HCV most conserved - Answers -5'UTR
Mantle (MCL) - Answers -t (11;14)
, Histones - Answers -Ionic
HER2 overexpression - Answers -Treatment with Traustmaub
What is FLT3 associated with? - Answers -AML
HFE - Answers -C282Y, H63D, S65C
Acute Promylocytic Leukemia (APL) - Answers -t(15;17)
Something about getting quality of RNA? Choices were all 3 letter acronym with R, like
RFN, RIN - Answers -The RNA integrity number (RIN) is an algorithm for assigning
integrity values to RNA measurements. The integrity of RNA is a major concern for
gene expression studies and traditionally has been evaluated using the 28S to 18S
rRNA ratio, a method that has been shown to be inconsistent.
Amplification method with polymerase creating a synthetic dsDNA - Answers -TBC
Identical twins for organ donor what should you do? - Answers -continue with assay, tell
supervisor, something about parents?
Why does NGS use fluorescence over spectrometry? - Answers -Fluorescence is more
sensitive because of the different ways of measuring absorbance and fluorescence.
Light absorbance is measured as the difference in intensity between light passing
through the reference and the sample. In fluorescence the intensity is measured
directly, without comparison with a reference beam.
Similarities between NGS and CE/Sanger - Answers -•DNA polymerase incorporates
fluorescently labeled dNTPs into a DNA template
•The newly incorporated nucleotides are identified by fluorophore excitation.
MALDI-Tof - Answers -Matrix-Assisted Laser Desorption/Ionization-Time Of Flight
(MALDI-TOF) Mass Spectrometry (MS) is a common method used for quality control
(QC) of oligonucleotides. MALDI-TOF instrumentation is suitable for use in high-
throughput industrial QC settings and can resolve molecules in the size range of
oligonucleotides
Heteroduplex - Answers -are dsDNA complexes - consisting of two partly mismatched
polynucleotide strands derived from two different parent molecules. (look up Denaturing
HPLC)
Benefits of NGS Output - Answers -Reads are relatively short, but so much more is
produced (100K-1 Billion) creating a huge amount of data for very little cost
