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MOLBIO Flashcards

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1
Q

Detection and characterization of nucleic acids and the products they encode for the purpose of providing clinical information

A

Molecular Diagnostics

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2
Q

3 Major Steps/Process in Molecular Diagnostics Lab

A

1) DNA/RNA Extraction and Purification
2) Amplification
3) Detection and Characterization

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3
Q

DNA or RNA is released from cells and separated from unwanted cellular debris such as proteins, lipids and carbohydrates

A

DNA/RNA Extraction and Purification

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4
Q

Billions of copies are produced from small amounts of target DNA/RNA from a sample

A

Amplification

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5
Q

Increases analytical sensitivity of molecular diagnostic tests

A

Amplification

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6
Q

Most common amplification

A

PCR

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7
Q

The presence,amount,size and specific sequences can be determined using various technologies such as spectrophotometry, fluorometry, electrophoresis and hybridization

A

Detection and Characterization

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8
Q

Functional unit within a larger nucleic acid molecule that code for a specific protein or function

A

Gene

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9
Q

Gene are made up of either

A

DNA or RNA

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10
Q

The collection of genes in a cell or organism

A

Genome

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11
Q

Genomes are organized into

A

Chromosomes

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12
Q

Single chromosomes is considered

A

1 DNA molecule

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13
Q

May be found in the mitochondria of eukaryotic cells,as plasmids in bacteria or in viral genomes

A

Double stranded DNA

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14
Q

DNA or RNA are made up of monomers called either

A

Nucleosides or Nucleotides

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15
Q

3 components of nucleotides

A

5 carbon sugar
Phosphate group
Nitrogenous base

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16
Q

5 carbon sugar can either be

A

Ribose or Deoxyribose

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17
Q

Phosphate group are esterified at what position

A

5’

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18
Q

Nitrogenous base are found in what position

A

1’

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19
Q

Nitrogenous base of RNA

A

A,C,G,U

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20
Q

Nitrogenous base of DNA

A

A,C,G,T

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21
Q

Missing OH group in Carbon 2

RNA/DNA

A

DNA

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22
Q

OH group is attach to what position

A

3’

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23
Q

Composed of Base + Sugar

A

Nucleoside

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24
Q

Composed of Base + Sugar+ Phosphate group

A

Nucleotide

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25
Form the base for the information stored in each gene
Nitrogenous bases
26
Nitrogenous bases may ne classified structurally either as
Purine or Pyrimide
27
Number of rings in pyrimidine
1
28
Number of rings in purine
2
29
Examples of Pyrimidine
Thymine/Uracil Cytosine
30
Examples of Purine
Adenine Guanine
31
Pyrimidine of RNA
Uracil Cytosine
32
Nitrogenous bases form complementary pairing via
Hydrogen bonds
33
Thymine is pair with
Adenine
34
Uracil is pair with
Adenine
35
Cytosine is pair with
Guanine
36
Thymine/Uracil pairs with Adenine forming how many hydrogen bonds
2
37
Cytosine-Guanine base pairs forming how many hydrogen bonds
3
38
Which nitrogenous base pair has stronger bonds
Cytosine-Guanine
39
Major factor in determining the melting point of DNA
Cytosine-Guanine base pair
40
Carries the genetic blueprint of an organism
DNA
41
DNA is usually found in
Nucleus or mitochondria
42
DNA is a: Double/Single strand
Double strand
43
Strands run in an
Anti-parallel fashion
44
DNA size or length is expressed according to the number of ______ or ______
Nucleotides Base pairs
45
Which is more stable? RNA/DNA
DNA
46
Why does DNA more stable
1) Double stranded and protected from mutagens 2) Less prone to hydrolysis reaction due to absent of hydroxyl group
47
Direction of DNA replication
5' to 3' direction
48
Template of DNA replication
3' to 5'
49
Primers are removed by
DNA polymerase I
50
Hydroxyl group is attach to what position
3'
51
Phosphate group is attach to what position
5'
52
Replication is usually
Bidirectional
53
Where is the origin of replication
At the center of a replication bubble
54
Each replication bubble have a _____ and _____ strand
Leading Lagging
55
Faster to synthesize because it has fewer step involve Lagging/ Leading?
Leading
56
Major enzymes in DNA replication
DNA polymerase III DNA polymerase I Helicase Primase Ligase
57
First step in DNA replication
Primer Annealing
58
DNA polymerase can only attach new nucleotides if there is 5' or 3' end
3' end
59
Site of growth of new DNA strands
3' end
60
It produce the complementary strand/fragments
DNA polymerase III
61
What type of DNA polymerase is for bacteria
DNA polymerase II
62
Fills the nucleotides of okazaki fragments
DNA polymerase I
63
Unwind/unzipped double stranded DNA
Helicase
64
Synthesize the primers
Primase
65
Attaching the okazaki fragments to form a single molecule
Ligase
66
Responsible for converting the information in DNA into proteins
RNA
67
3 types of RNA
mRNA rRNA tRNA
68
Not stable, prone to hydrolysis, susceptible to mutation
RNA
69
RNA is : Double or single strand
Typically Single strand
70
RNA is synthesized by what enzyme
RNA polymerase
71
Short RNA strand to which DNA polymerase can attach the first nucleotide during DNA strand synthesis
Primers
72
Later removed and replaced by DNA
Primers
73
Converts DNA into mRNA
Transcription
74
Decodes mRNA into proteins such as amino acids
Translation
75
4 Samples in Nucleic acid detection
Cell suspension Tissue sample Microbial cultures Cell free samples (ex: serum)
76
Contains more chromosomal DNA RBC/ WBC?
WBC
77
Blood, CSF, feces, urine,exudates,transudates, swabs
Cell suspension
78
More invasive sample in nucleic acid detection
Tissue samples
79
May be fresh,frozen,fixed,human or animal(parasite) tissue
Tissue samples
80
Fixatives used in tissue sample for nucleic acid detection which are considered least damaging
10% buffered formalin Acetone Alcohol
81
Cultures of bacteria, fungi, or virus may be analyzed for identification and characterization
Microbial cultures
82
May still contain enough DNA or RNA to be detectable when amplification is employed
Cell free samples (ex: Serum)
83
Mitochondrial DNA can be isolated from: Nucleated/ Non-nucleated cells
Non-nucleated cells
84
mRNA are compose of
Exons Introns
85
Contains instructions of making DNA/structural coding
Exons
86
Nonstructural coding
Introns
87
6 Types of Release/Lysis Methods
1) Mechanical method (grinding/mincing) 2) Heating/Boiling 3) Strong alkali (NaOH) 4) Sonication 5) Enzyme/Protease 6) Detergent (SDS, Triton X)
88
Lysis method that use mortar and pestle
Mechanical method
89
Lysis method that is most convenient and cheap method
Heating/Boiling
90
Lysis method that use reagent kit and sodium hydroxide
Strong alkali (NaOH)
91
Lysis method that se soundwaves
Sonication
92
6 DNA Purification/Isolation method
Centrifugation Organic Isolation Inorganic Isolation Solid-Phase Isolation Chelating Ion Exchange Resin Magnetic Beads
93
Most efficient but toxic isolation method
Organic Isolation (Phenol/Chloroform Isoamyl Method)
94
Primary reagent used in Organic Isolation that dissolves out lipids and proteins
Phenol Chloroform
95
In organic isolation, DNA will be found in the
Upper aqueous layer
96
Use to precipitate out nucleic acid in organic isolation
Alcohol
97
In organic isolation, after centrifuge,DNA/RNA will form a
pellet
98
Uses low pH and high salt conditions to precipitate out proteins,leaving DNA in solution
Inorganic isolation(salting-out)
99
In Inorganic isolation,DNA is later precipitated using
Alcohol
100
Inorganic isolation is AKA
Salting-out
101
Most common use DNA isolation
Solid-Phase Isolation
102
Solid-Phase Isolation is based from?
Silica
103
Silica-based matrices in the form of beads or colums are used to bind DNA
Solid-phase isolation
104
To bind the nucleic acids to the silica matric, you can use
Chaotropic salts (guanine hydrochloride)
105
Cause DNA to bind to silica by weakening H bind with water and forming cation bridges
Chaotropic salts
106
In solid-phase isolation, DNA is eluted from the silica matric using a
Low salt buffer Deionized water
107
Major steps in Solid-Phase Isolation
Lysis Binding Washing Elution
108
A suspension of 5-10% chelex resin beads is mixed woth the specimen then boiled for lysis
Chelating Ion Exchange Resin (Chelex) Isolation
109
How many percent of chelex resin beads suspension is needed
5-10%
110
DNA are made to bind to magnetic beads coated with silicone dioxide
Magnetic Beads
111
DNA are made to bind to magnetic beads coated with
silicone dioxide
112
In magnetic bead, it is use to facilitate binding
Strong salts Alcohol
113
Magnetic particles are added to sample and bind to target molecule Bind/ Wash/ Elute?
Bind
114
Magnetic beads are captured and remainder of sample is washed away Bind/ Wash/ Elute?
Wash
115
Target molecule is released from magnetic particles Bind/ Wash/ Elute?
Elute
116

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