1.

Paternity Testing

· pC frequency of C allele in population

2. Common allele vs rare allele
3. Application of Human Identification (HI)? Why do we use chromosome in HI?
Y-chromosome:
● Application:

● Methods:
4.

5. Advantage and disadvantage of molecular biotech in microorganism detection

6. Huntington disease and V-leiden disease detection

Hungtinton’s disease detection V-Leiden disease way of detection

 - The genetic disorder is caused if - Factor V Leiden is a variant
they have just one disease-causing copy of (mutated form) of human factor V gene
a gene (Autosomal-dominant single gene F5 (one of several substances that helps
disorder). blood clot)

- Occurs in every generation. - Factor V variant can not be easily

degraded by activated Protein C- natural
Detection: simple PCR-based assays to anticoagulant àincrease in blood clotting -
determine the presence of the mutant allele thrombophilic phenotype.

+ The CAG repeat is amplified using - Mutation pattern : a SNP located in

primers HD1-short and HD3-3′. HD1-short exon 10 (substitution G to A <-> arginine
is the standard PCR primer developed for to glutamine).
this assay except that the 3′-most base was
Way of detection: PCR-RFLP PCR-
removed to avoid loss of annealing in those restriction fragment length polymorphism
cases with a C-to-G polymorphism of the (RFLP)-based analysis- a popular
base immediately preceding the repeat. technique can exploit SNPs with
+ The CCG/CCT repeat is amplified using inexpensiveness and lack of requirement
HD3-5′ and HD2. The entire repeat region for advanced instruments.
is amplified using HD1-short and HD2 or, to
+ The R506Q substitution G àA (exon 10
maximize independence of assays, primers
- F5 gene).
HU4 and HU3.
+ DNA mutation destroys MnlI restriction
+ PCR product size evaluated with enzyme site.
polyacrylamide gel electrophoresis. The
product length is detected by use of a + Amplicon when cut with Mnl I, will
radiolabeled primer to generate a yield :
radiolabeled product, a radiolabeled (CAG) n
or (CTG)n oligonucleotide to probe a *3 fragments in normal DNA (+/+)
Southern blot of the gel, or a fluorescently
tagged primer to generate fluorescent *2 products in homozygous (m/m).
products detected by an automated
genotyping system.
*A combination in heterozygous
specimen (+/m)
+ Control samples of known repeat length
are essential, especially in interpreting
borderline expansions, because repeat
expansion may slightly alter the
electrophoretic mobility relative to standard
size markers

7. Principle of clonality. White blood cancer cell detect

8. Southern blot using for gene rearrangement

9. Method for HLA typing

10. Cell-free DNA for Down syndrome detection?
● Definition: Cell-free DNA screening is a test that can determine if a woman has a higher
chance of having a fetus with Down syndrome (trisomy 21)
● Đối tượng:
Phụ nữ ngoài 35
Phụ nữ đã từng có con mắc bệnh Down
Other woman with lower-risk: should use the traditional technique to diagnose for more
disorder.
● Exception: Not accurate in women with twins, and cannot be used in women with triplets.
● Accuracy: More than 99 percent of Down syndrome pregnancies. If woman is high risk of
Down syndrome, she can choose to do further test to make sure.
● Sampling: woman's blood is taken after 10 weeks of pregnancy.
Measures the small fragments of fetal DNA in the mother's blood -> determine the chance of a
chromosome problem based on the relative amount of DNA from chromosomes 21, 18, 13 and
the sex chromosomes.
● Methods: 4 methods
WGS: Sequence both mom and baby DNA fragment. Identifying and counting large number of
DNA fragment then compare to the reference. If there is a relative excess or deficit -> fetal
aneuploidy
SNP: SNP sequencing, using blood sample:
+ mother plasma -> SNP of fetus and mom
+ mother WBC -> SNP of mom
Then analyze and compare them -> detect trisomy
Targeted capture enrichment methodology: CfDNA is counted by NGS and then align on
targeted region of human genome.
Microarray-based targeted methodology: Using DANSR assay

All methodologies analyzed the total (maternal and fetal) cfDNA. Minimum fetal fraction (ff) is
4%.