Diagnostic testing

Question 1

ASO

Answer 1

• Small and specific (15bp)
• Point mutation

Disease:
1) Cystic Fibrosis
2) Hemochoromatosis
3 SICKLE CELL

Question 2

ASO PCR

Answer 2

-Identifies single nucleotide mutation or destruction of restriction site

Triple repeats Disorder:

1) Huntington (CAG
2) Fragile X (CGG)
3) Myotonic Dystrophy (CTG)

Question 3

RFLP

Answer 3

-mutations and restriction sites

Diseases:
1) SICKLE CELL

Question 4

Linkage Analysis

Answer 4

-Indirect detection
-Determine carrier status
Buzzword: Marker

Diseases:

1) Allelic heterogeneity
2) Cystic fibrosis
3) Hemochromatosis
4) Neurofbromatosis Type I

Question 5

Sky Fish

Answer 5

1) Translocation (EX: Cancer)

2) Paints chromosomes different colors so can’t observe deletions

Question 6

Array CGH

Answer 6

• Multiple colors
• Tests for duplications and deletions
• Uses millions of probes
• ENTIRE GENOME
• Red =deletions
• green= duplications
• Misses translocations
• Copy number variant

Diseases:
1) FIRST TIER TEST FOR CHILDREN W DEVELOPMENTAL DELAY AND MENTAL IMPAIRMENT

2) FIRST TIER TEST WHEN YOU DON’T KNOW WHAT GENE IT IS (genetic diagnosis)
3) Trisomy 18
4) Cancer

Question 7

SNP

Answer 7

• Point mutations
• Polymorphism
• Million probes

Common testing:
1) Paternity test

2) Haploid analysis (Imprinting disorder) (Ex: Prader willie and angelman)

Question 8

PCR

Answer 8

Length of DNA
Restriction enzyme

Disease:

• TRIPLE REPEAT
• Cystic fibrosis

Question 9

Next Generation

Answer 9

• Whole genome sequencing
• Whole exome sequencing
• Single gene analysis2

-MASSIVE PARALLEL SEQUENCING

Question 10

Gel Electrophoresis

Answer 10

Keywords:
"a well"
"gel"
"Agrose"
"UV"

Question 11

Southern Blots

Answer 11

DNA

-TRIPLE REPEATS

DNA is purified from a cell and then cut and then with restriction enzymes and separated by gel electrophoresis

Looking for deletions and insertions

Question 12

Northern Blots

Answer 12

mRNA

Detects amount and size of the particular mRNA of interest

Question 13

Western Blots

Answer 13

Proteins (Exome/exons, protein coding)

Question 14

cDNA Microarray

Answer 14

• Isolates RNA
• Comparison of gene expression between cell or tissue sample

-Red= over-expression
-Green= under-expression
yellow=equal abundance

Question 15

Techniques that detect only what is asked for

Answer 15

1) ASO blot
2) PCR
3) Southern
4) FISH
5) Sanger

Question 16

FISH

Answer 16

• Higher resolution
• Cannot detect small deletions
• Need to know what you are looking for
• Hybridizes ssDNA sequences to Metaphase chromosome

Diseases:

1) DE GEORGE (22q11.2)
2) PRADER WILLI/ANGELMAN CHIPS

Symptoms De George: 
-Cardiac defects
-Abnormal feces
-Thymic
-Cleft Palate
-Hypocalemia
(Chromosome 22)

Question 17

Rapid FISH

Answer 17

• Pressed for time

- Done in interphase