3SQ1 Flashcards
The following image is called a _______, the chromosomes are in their ______ form
Karyogram
Picture of Karyogram (di ko malagay ei)
Karyogram; haploid
T/F: The previous figure is an example of symmetric karyotype.
F ; asymmetric
Contains: Glutathione, Vit B12, NaHCO3
For which part of the process of karyotyping is this used for?
Cell culture
BONUS: Spell out RPMI
Roswell Park Memorial Institute
Select all that apply
Which other reagents and equipment are needed for your answer in 3 (Cell culture)?
Fetal BSA, PHA
What is the role of choice B (PHA) in karyotyping preparation?
Cell division factor
BONUS: The blood to be used in lymphocyte based karyotyping must have ______ as anticoagulant, and therefore the tube top should be ________ colored.
Sodium Heparin, green
The main principle behind ________ method is to arrest cells at _________ by disruption of the ________ via the use of drug
Colcemid, metaphase, microtubules
Which reagent in the karyotyping procedure cause the following to retrieve chromosomes?
Sodium citrate 0.8%
BONUS: ________ exposure to colcemid or the use of high concentrations increases the proportion of chromosomes at late metaphase, resulting in shortening of the chromosomes. A ______ exposure with a high concentration of colcemid reduces the total yield of metaphases.
Long, short
BONUS: This (Bromouridine) is used to arrest the cells at ___-phase to increase the total yield of metaphase chromosomes.
Interphase
Identify the ingredients and the ratio of ingredients in Carnoy’s fixative
Ethanol; Ch3Cl; 95% HOAc (6:3:1)
For which part of the process is the inverted microscope used?
Step where rubber policeman is used
Cell culture; to spread the cells in a culture
Together with the ingredient seen in ________ (reagent), this equipment (Carbon Dioxide incubator) is used to provide proper temperature and ________ to the cell culture.
RPMI, pH
Which is not properly matched?
Final fixation - Room temp (Cold)
G-banding Karyotype
- Uses eosin Y and azure B
- The positive G-bands, which are the dark bands, are hydrophobic regions
- Lighter stained regions are disulfide-rich
R-banding karyotype
- Produces positive bands that correspond to the negative bands in G-banding
- Is produced by incubating the chromosomes in an ionic solution at a high temperatures (~87C) followed by staining with Giemsa
C-banding karyotype
- Technique involves hot saline incubation, and alkali treatment
Q-banding karyotype
- Dye binds by intercalation
- Amino group at position 2 of the Guanine bases of the DNA quenches the fluorescence of quinacrine
- AT-rich regions of the chromosomes to fluoresce more brightly than the GC-rich regions
NOR-banding karyotype
- Acetic acid is used like a stop bath to end the staining process
BONUS: Identify the karyotype staining with the following advantages:
- Identification of bivalents at diakinesis using both centromere position
- Paternity testing
- Gene mapping
C-banding
Identify the sex of the human using the previous karyotype:
Male
Trisomy 21 Karyotype
Down Syndrome
XXY Karyotype
Klinefelter Syndrome
Something sa chromosome 15
Angelman syndrome daw
5p- deletion
Cri-du-chat
Which is correct about amniocentesis?
- Test specifically looks at Chromosomes 21, 18, 13, X and Y
- Can be used in classical karyotyping
T/F: A prenatal test checks cells from the placenta (which are identical to cells from the fetus) to see if they have a chromosomal abnormality. The test can be done from weeks 10 to 13 in a woman’s pregnancy.
T
T/F: A transmission electro microscope is required in order to visualize the patient’s karyotype. The electron micrograph of the patient’s karyogram will then be compared against the karyogram from a donor sample without mutation.
F
Not true of FISH
- Looks for specific protein antigens in tissue
- Can detect point mutations
Identify part of FISH
Probe DNA
Which is the sequence of previously seen technique?
Sample preparation > denaturation > hybridization > washing > visualization
Select all that is true to answer in 26 (Probe DNA)
- Length is 20-1000bps
- Can be RNA
- Can be DNA analogs
- Can be tagged with nonfluorescent molecule first (Indirect Fluorescence)
Select all that applies to target DNA
- Patient’s own DNA
Short stouted chromosomes
Metaphase
basta yung 31 may red at green image
Interphase
Which is correlated with 31 (Interphase)?
- Can be used for cytospin and FFPE
- Can be used for de novo changes
BONUS: Spell out FFPE
Formalin Fixed Paraffin Embedded
Choose the letter of Indirect Fluorescence FISH
B (Streptavidin)
What is the missing hapten in your answer to 33 (Streptavidin)?
Biotin
BONUS: Egg ___ is a source of your answer to Biotin
yolk
FITC
Green
Rhodamine
Red
DAPI
Blue
BONUS: A cytotechnician was asked to prepare a bone marrow aspirate for FISH analysis. After mixing the probe and the sample, he incubated the mixture and focused the preparation on the microscope without washing. Which of the following would be the most likely result?
False positive
Lower temperatures for denaturation
Formamide
50-70-90-100; ascending
Ethanol
Which kind of FISH is this (multiple colors)
Locus-specific probes
Centromeric staining sa FISH
Alphoid probes
Subtelomeric probes; This technique in the figure can detect:
- Loss of tip regions of linear chromosome
Mband Analysis
Intrachromosomal structural change
Multiplex FISH
Color-tagging for cytogenetics of pre-B leukemia
FIBER Fish
High resolution mapping of telomeric chromosomes
BONUS: Identify the technique
Dual Fusion
BONUS: Which cancer is correlated with HER2/neu gene
Breast cancer
Select all that is true of the technique (CGH)
- Used for detection of copy number loss/gain
- Used for detection of whole genome not transcriptome
- CGH
Which is not correlated with the previous figure (CMA)
- Uses CCD as light source
Which chromosomal abeeration can be represented by the red line? (Gene deletion)
CFTR
This step:
Stringent washing
Identify this microscope used in FISH
Fluorescence
BONUS: ______ is prepared exclusively from male human placental DNA by shearing, denaturing, and reannealing under conditions that enrich repetitive elements
Cot 1
BONUS: In Carnoy’s fixative, which substance is used to coagulate nucleoproteins and cause swelling of the cells. This fixative penetrates the cells rapidly and preserves the chromosome structure.
Acetic Acid
