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Hybridization Techniques Flashcards

1
Q

It is the phenomenon in which DNA/RNA/Protein anneal to complementary DNA/RNA/Protein

A

Hybridization techniques

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2
Q

It recognizes specific base sequences and cuts the DNA. It is also considered an endonuclease that is naturally found in bacterial cells.

A

Restriction Enzymes

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3
Q

What are the 4 Classifications of Restriction Enzymes?

A

Type I - IV

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4
Q

It contains both nuclease and methylase activity and can recognize specific sequences and cut DNA at a NON-SPECIFIC SITE > LESS THAN 1000 BP AWAY

A

Type I

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5
Q

It is commonly used in the laboratory and is considered unfunctional that can recognize (blank) and cut within the palindrome

A

Type II; Palindromic sequence

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6
Q

It is bifunctional and can recognize specific 5-7 bp sequences and cut 24-24 bp down

A

Type III

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7
Q

What are the different blotting techniques?

A

SNoW
Southern Blots
Nothern Blots
Western Blots

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8
Q

Macroarray and Microarray are types of?

A

Genomic Array Technology

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9
Q

It is the binding of complementary sequences and the formation of a duplex between 2 complementary nucleotide sequences

A

Hybridization

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10
Q

Probes are used for?

A

Detection of specific sequence

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11
Q

What is a primer?

A

It is the starting point of Replication (RNA) ; in vitro = Amplification (DNA)

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12
Q

It is the process of transferring sample nucleic acids on a solid support and is widely used for the identification of desired DNA, RNA fragments, and proteins from thousands of molecules.

A

Blotting techniques

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13
Q

Southern blot detects?

A

DNA

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14
Q

(blank) is isolated and cut with restriction enzymes

A

DNA

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15
Q

DNA fragments are separated by?

A

Gel electrophoresis

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16
Q

It is the process of removing purine bases that has Larger DNA fragments (>500 bp)

A

Depurination

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17
Q

The gel is treated with (blank) on Denaturation

A

NaOH

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18
Q

To move the DNA from the gel to a membrane substrate from probing

A

Blotting (Transfer)

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19
Q

Give 2 membrane types

A

Pure nitrocellulose and Reinforced nitrocellulose

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20
Q

Give 3 transfer methods

A

Capillary transfer, Electrophoretic transfer, and Vacuum transfer

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21
Q

It is driven by the capillary movement of a buffer from the soaked paper to the dry paper.

A

Capillary transfer

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22
Q

driven by the capillary movement of a buffer from the soaked paper to the dry paper, (blank) moves from the gel to the membrane. the (blank) will adhere to the membrane, which will be subsequently exposed to the probe.

A

denatured DNA; DNA

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23
Q

This system uses electric current to move the DNA transversely through the get to the membrane. This type of transfer is used mostly for small fragments or proteins.

A

Electrophoretic System/Transfer

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24
Q

This system uses suction and buffer recirculation to move the DNA out of the gel and onto the membrane.

A

Vacuum transfer

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25
It is generally faster than capillary transfer for (blank) DNA fragments. However, unlike capillary transfer, (blank) requires specialized equipment.
large; vacuum transfer
26
A process to prevent the transferred DNA fragments from washing away or moving on the membrane
Immobilization of DNA
27
Bake the membrane in a (blank) oven at what degrees with time?
Vacuum; 80*C - 30 to 60 mins
28
This prevents the probe from binding to nonspecific sites on the membrane surface
Prehybridization
29
True or False: Buffer does contain probe in Prehybridization
FALSE Buffer does NOT contain probe in Prehybridization
30
True or False: Before hybridization, the excess probe is washed from the membrane
FALSE AFTER hybridization, the excess probe is washed from the membrane
31
It is when the pattern of hybridization is visualized
Detection
32
What are the two ways to detect patterns of hybridization?
Xray film and Chromogenic detection method
33
It is the modification of the Southern blot technique
Northern blots
34
This blotting technique is designed and developed by James Alwine to investigate RNA
Northern blots
35
This blotting technique is another modification of the Souther blot that immobilized proteins
Western Blots
36
What is the target cell of Southern blot and its probe?
DNA ; Nucleic Acid
37
What is the target cell of Northern blot and its probe?
RNA ; Nucleic Acid
38
What is the target cell of Western blot and its probe?
Protein ; Protein
39
What is the target cell of Southwestern blot and its probe?
Protein ; DNA
40
What is the purpose of Southern blot? A. Gene Structure B. Protein processing, gene expression C. Transcript structure, processing, gene expression D. DNA binding proteins, gene regulation
A. Gene Structure
41
What is the purpose of Northern Blot? A. Gene Structure B. Protein processing, gene expression C. Transcript structure, processing, gene expression D. DNA binding proteins, gene regulation
C. Transcript structure, processing, gene expression
42
What is the purpose of Western Blot? A. Gene Structure B. Protein processing, gene expression C. Transcript structure, processing, gene expression D. DNA binding proteins, gene regulation
B. Protein processing, gene expression
43
What is the purpose of Southwestern Blot? A. Gene Structure B. Protein processing, gene expression C. Transcript structure, processing, gene expression D. DNA binding proteins, gene regulation
D. DNA binding proteins, gene regulation
44
These are small fragments of nucleic acid that are complementary to a part of the DNA or RNA of the organism we are searching for
Probes
45
What are the two types of probe labeling?
Radioactively and Fluorescently
46
What are the two types of fluorescent labeling?
Biotin and Digoxygenin
47
It is a more specific type of probe that is expensive and difficult to synthesize which may require reaction times of hours to hybridize
Long probes (500-5000 bp)
48
It hybridizes at very high rates and is less specific
Shorter probes (<500 bp)
49
What are the factors affecting stringency?
Temperature, salt concentration of the hybridization buffer, and concentration of denaturant
50
Tm = 4 x number of GC pairs + 2 x number of AT pairs This formula is used calculate the?
Melting temperature
51
The determination of the size of the target is not required and is deposited directly on the membrane
Dot/Slot Blot
52
The target is deposited in a circle or dot
Dot/Slot Blots
53
The target is deposited in an oblong bar
Slot Blots
54
The targets can be screened simultaneously in a very small area by miniaturing the deposition of droplets
Microarrays
55
It is used to measure mRNA expression and is more sensitive than Northern blotting. It is also used to detect sequence changes in DNA or mutational analysis.
Solution hybridization
56
Fluorescent In Situ Hybridization is also known as?
In Situ Hybridization or ISH
57
It is used to detect and localize the presence or absence of specific DNA sequences on chromosomes
ISH
58
What are the requirements for FISH?
Sample, Fluorescent Probe and Fluorescence Microscope
59
The label is bound to the probe
Direct labelling
60
It requires an additional step before detection. The probe was detected using antibodies conjugated to labels.
Indirect labeling
61
A fluorescent probe attaches to the target sequence during hybridization and is visualized through a fluorescence microscope
Visualization

Molecular Biology (1 decks)

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