Laboratory techniques Flashcards
Temperature of denaturation in PCR
96 degree c
Gice examples for PCR use
Neonatal HIV
herpes encephalitis
Give the temperature for annealing
55 degree C
Give the temperature for Elongation
72 degree c
Name the substrates for denaturation
DNA template
DNA primers
Taq Polymerase
dNTP
Average time for one PCR
10 minutes
From which organism is Taq polymerase acquired
Thermus Aquaticus
After 3 cycles the amount of DNA in PCR is
8
2^n
Variation of PCR used in RNA viruses
RTPCR
Which swab is taken in case of COVID 19
Nasopharyngeal swab
Which RNA virus was tested in COVID 19
SARS COV 2
What is the full form of CRISPR
Clustered Random Interspace Short Palindromic Repeats
What all is required in CRISPR technology
Cas9 (endonuclease)
Guide RNA
Template DNA
What are the 2 editing methods in CRISPR
- Non homologous end joining in frameshift mutation
- Homologous joining by Donor DNA
1- knock out 2- knock in
Device used for PCR
Thermal Cycler
How to identify Fragile X Syndrome with the help of Southern Blot
Identifying the no of CGG repeats in FMR 1
Name the electrophorized substance in
Southern blotting
Northern
Western
Southwestern
Southern- DNA
Northern - RNA
Western- Protein
Southwestern- DNA binding protein
SNoW DRoP
What all can be found out by flow cytometry
Size
Granularity
Protein expression
What all is measured in flow cytometry
Light scatter
Intensity of fluorescence
What diseases is flow cytometry used for
Hematological abnormalities (leukemia ; paroxysmal nocturnal hemoglobin etc.)
Immunodeficiencies (CD4+ cell count in HIV)
Name the plots used for one and two measure in flow cytometry
one- histogram
two - Scatter plot
In microarrays ______ are arranged on a grid on a glass or silicon chip
DNA oligonucleotides
What does the ratio of fluorescence signal tell us in microarrays
Relative amount of hybridizing nucleic acids in those samples
Use of microarray
compare relative transcription of gene in 2 RNA samples
Detection of SNP and CNV used in-
Genotyping
Clinical genetic testing
forensic analysis
cancer mutation
genetic linkage analysis
Which sample is used in ELISA
Blood
Urine
Sputum
Stool
Blood
Which is more specific, western blot or ELISA
Western blot
How is direct ELISA different from indirect ELISA
Indirect Elisa makes use of a secondary antibody attached to a primary antibody
What drug is used in karyotyping to halt chromosomes at ______ stage
Colchicine; metaphase
Chromosomes are stained according to ______
Morphology
Size
Arm Length Ratio
Banding pattern
What all samples can be used in Karyotyping
Blood
Bone marrow
Amniotic fluid
Placental tissue
What are the 3 type of chromosomal abnormalities that can be seen in fluorescence in situ hybridization
Microdeletion- no fluorescence
Translocation- Fluorescence signal from one gene in one chromosome to another chromosome
Duplication- a second copy of chromosome
Give an example of translocation
Fluoroscence signal from ABL gene on chromosome 9 goes near to BCR gene on chromosome 15
What is molecular cloning used for
Production of human protein in bacteria eg insulin
Steps for molecular cloning
read in usmle pg 53
______ insertion (random/targeted) has _______ expression (conditional/constitutive)
Random- constitutive
Targeted- conditional
Process by which small non coding RNA molecules target mRNA to inhibit gene expression is called
RNA interference
_____ are used for a broad targeting of gene suppression
Micro RNA
blocks translation
siRNA are derives form
Exogenous dsRNA source like virus
_____ are used for a specific targeting of gene suppression
siRNA
it is chemically synthesized for gene knockdown experiments
What protein help in putting environmental stress on bacteria to incorporate the plasmid
Heat Shock Protein
What is Polymorphic Marker
Marks out specific spots to identify polymorphism
After a DNA sequence is inserted into a plasmid for cloning, the resultant molecule is known as a _______plasmid.
Recombinant
The process that involves introducing a plasmid into bacteria is called _______.
Transformation
To isolate a specific DNA region in the polymerase chain reaction, researchers synthesize _________that act as primers
Oligonucleotides
Other than the gene of interest, plasmids for DNA cloning contain an _______ gene which confers antibiotic protection to the bacteria with the plasmid.
antibiotic resistance
When analyzing a gel, bands of macromolecules are compared to a standard ladder to determine (size/charge) .
Size
