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Cellular biochemistry > Laboratory techniques > Flashcards

Laboratory techniques Flashcards

1
Q

Temperature of denaturation in PCR

A

96 degree c

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2
Q

Gice examples for PCR use

A

Neonatal HIV
herpes encephalitis

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3
Q

Give the temperature for annealing

A

55 degree C

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4
Q

Give the temperature for Elongation

A

72 degree c

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5
Q

Name the substrates for denaturation

A

DNA template
DNA primers
Taq Polymerase
dNTP

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6
Q

Average time for one PCR

A

10 minutes

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7
Q

From which organism is Taq polymerase acquired

A

Thermus Aquaticus

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8
Q

After 3 cycles the amount of DNA in PCR is

A

8

2^n

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9
Q

Variation of PCR used in RNA viruses

A

RTPCR

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10
Q

Which swab is taken in case of COVID 19

A

Nasopharyngeal swab

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11
Q

Which RNA virus was tested in COVID 19

A

SARS COV 2

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12
Q

What is the full form of CRISPR

A

Clustered Random Interspace Short Palindromic Repeats

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13
Q

What all is required in CRISPR technology

A

Cas9 (endonuclease)
Guide RNA
Template DNA

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14
Q

What are the 2 editing methods in CRISPR

A
  1. Non homologous end joining in frameshift mutation
  2. Homologous joining by Donor DNA

1- knock out 2- knock in

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15
Q

Device used for PCR

A

Thermal Cycler

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16
Q

How to identify Fragile X Syndrome with the help of Southern Blot

A

Identifying the no of CGG repeats in FMR 1

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17
Q

Name the electrophorized substance in
Southern blotting
Northern
Western
Southwestern

A

Southern- DNA
Northern - RNA
Western- Protein
Southwestern- DNA binding protein

SNoW DRoP

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18
Q

What all can be found out by flow cytometry

A

Size
Granularity
Protein expression

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19
Q

What all is measured in flow cytometry

A

Light scatter
Intensity of fluorescence

20
Q

What diseases is flow cytometry used for

A

Hematological abnormalities (leukemia ; paroxysmal nocturnal hemoglobin etc.)
Immunodeficiencies (CD4+ cell count in HIV)

21
Q

Name the plots used for one and two measure in flow cytometry

A

one- histogram
two - Scatter plot

22
Q

In microarrays ______ are arranged on a grid on a glass or silicon chip

A

DNA oligonucleotides

23
Q

What does the ratio of fluorescence signal tell us in microarrays

A

Relative amount of hybridizing nucleic acids in those samples

24
Q

Use of microarray

A

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

25
Q

Which sample is used in ELISA
Blood
Urine
Sputum
Stool

A

Blood

26
Q

Which is more specific, western blot or ELISA

A

Western blot

27
Q

How is direct ELISA different from indirect ELISA

A

Indirect Elisa makes use of a secondary antibody attached to a primary antibody

28
Q

What drug is used in karyotyping to halt chromosomes at ______ stage

A

Colchicine; metaphase

29
Q

Chromosomes are stained according to ______

A

Morphology
Size
Arm Length Ratio
Banding pattern

30
Q

What all samples can be used in Karyotyping

A

Blood
Bone marrow
Amniotic fluid
Placental tissue

31
Q

What are the 3 type of chromosomal abnormalities that can be seen in fluorescence in situ hybridization

A

Microdeletion- no fluorescence
Translocation- Fluorescence signal from one gene in one chromosome to another chromosome
Duplication- a second copy of chromosome

32
Q

Give an example of translocation

A

Fluoroscence signal from ABL gene on chromosome 9 goes near to BCR gene on chromosome 15

33
Q

What is molecular cloning used for

A

Production of human protein in bacteria eg insulin

34
Q

Steps for molecular cloning

A

read in usmle pg 53

35
Q

______ insertion (random/targeted) has _______ expression (conditional/constitutive)

A

Random- constitutive
Targeted- conditional

36
Q

Process by which small non coding RNA molecules target mRNA to inhibit gene expression is called

A

RNA interference

37
Q

_____ are used for a broad targeting of gene suppression

A

Micro RNA

blocks translation

38
Q

siRNA are derives form

A

Exogenous dsRNA source like virus

39
Q

_____ are used for a specific targeting of gene suppression

A

siRNA

it is chemically synthesized for gene knockdown experiments

40
Q

What protein help in putting environmental stress on bacteria to incorporate the plasmid

A

Heat Shock Protein

41
Q

What is Polymorphic Marker

A

Marks out specific spots to identify polymorphism

42
Q

After a DNA sequence is inserted into a plasmid for cloning, the resultant molecule is known as a _______plasmid.

A

Recombinant

43
Q

The process that involves introducing a plasmid into bacteria is called _______.

A

Transformation

44
Q

To isolate a specific DNA region in the polymerase chain reaction, researchers synthesize _________that act as primers

A

Oligonucleotides

45
Q

Other than the gene of interest, plasmids for DNA cloning contain an _______ gene which confers antibiotic protection to the bacteria with the plasmid.

A

antibiotic resistance

46
Q

When analyzing a gel, bands of macromolecules are compared to a standard ladder to determine (size/charge) .

A

Size

Cellular biochemistry (6 decks)

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