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MOLBIO Flashcards

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
Q

Form the base for the information stored in each gene

A

Nitrogenous bases

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26
Q

Nitrogenous bases may ne classified structurally either as

A

Purine or Pyrimide

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27
Q

Number of rings in pyrimidine

A

1

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28
Q

Number of rings in purine

A

2

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29
Q

Examples of Pyrimidine

A

Thymine/Uracil
Cytosine

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30
Q

Examples of Purine

A

Adenine
Guanine

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31
Q

Pyrimidine of RNA

A

Uracil
Cytosine

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32
Q

Nitrogenous bases form complementary pairing via

A

Hydrogen bonds

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33
Q

Thymine is pair with

A

Adenine

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34
Q

Uracil is pair with

A

Adenine

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35
Q

Cytosine is pair with

A

Guanine

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36
Q

Thymine/Uracil pairs with Adenine forming how many hydrogen bonds

A

2

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37
Q

Cytosine-Guanine base pairs forming how many hydrogen bonds

A

3

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38
Q

Which nitrogenous base pair has stronger bonds

A

Cytosine-Guanine

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39
Q

Major factor in determining the melting point of DNA

A

Cytosine-Guanine base pair

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40
Q

Carries the genetic blueprint of an organism

A

DNA

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41
Q

DNA is usually found in

A

Nucleus or mitochondria

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42
Q

DNA is a:

Double/Single strand

A

Double strand

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43
Q

Strands run in an

A

Anti-parallel fashion

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44
Q

DNA size or length is expressed according to the number of ______ or ______

A

Nucleotides
Base pairs

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45
Q

Which is more stable?

RNA/DNA

A

DNA

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46
Q

Why does DNA more stable

A

1) Double stranded and protected from mutagens
2) Less prone to hydrolysis reaction due to absent of hydroxyl group

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47
Q

Direction of DNA replication

A

5’ to 3’ direction

48
Q

Template of DNA replication

A

3’ to 5’

49
Q

Primers are removed by

A

DNA polymerase I

50
Q

Hydroxyl group is attach to what position

A

3’

51
Q

Phosphate group is attach to what position

A

5’

52
Q

Replication is usually

A

Bidirectional

53
Q

Where is the origin of replication

A

At the center of a replication bubble

54
Q

Each replication bubble have a _____ and _____ strand

A

Leading
Lagging

55
Q

Faster to synthesize because it has fewer step involve

Lagging/ Leading?

A

Leading

56
Q

Major enzymes in DNA replication

A

DNA polymerase III
DNA polymerase I
Helicase
Primase
Ligase

57
Q

First step in DNA replication

A

Primer Annealing

58
Q

DNA polymerase can only attach new nucleotides if there is

5’ or 3’ end

A

3’ end

59
Q

Site of growth of new DNA strands

A

3’ end

60
Q

It produce the complementary strand/fragments

A

DNA polymerase III

61
Q

What type of DNA polymerase is for bacteria

A

DNA polymerase II

62
Q

Fills the nucleotides of okazaki fragments

A

DNA polymerase I

63
Q

Unwind/unzipped double stranded DNA

A

Helicase

64
Q

Synthesize the primers

A

Primase

65
Q

Attaching the okazaki fragments to form a single molecule

A

Ligase

66
Q

Responsible for converting the information in DNA into proteins

A

RNA

67
Q

3 types of RNA

A

mRNA
rRNA
tRNA

68
Q

Not stable, prone to hydrolysis, susceptible to mutation

A

RNA

69
Q

RNA is :

Double or single strand

A

Typically Single strand

70
Q

RNA is synthesized by what enzyme

A

RNA polymerase

71
Q

Short RNA strand to which DNA polymerase can attach the first nucleotide during DNA strand synthesis

A

Primers

72
Q

Later removed and replaced by DNA

A

Primers

73
Q

Converts DNA into mRNA

A

Transcription

74
Q

Decodes mRNA into proteins such as amino acids

A

Translation

75
Q

4 Samples in Nucleic acid detection

A

Cell suspension
Tissue sample
Microbial cultures
Cell free samples (ex: serum)

76
Q

Contains more chromosomal DNA

RBC/ WBC?

A

WBC

77
Q

Blood, CSF, feces, urine,exudates,transudates, swabs

A

Cell suspension

78
Q

More invasive sample in nucleic acid detection

A

Tissue samples

79
Q

May be fresh,frozen,fixed,human or animal(parasite) tissue

A

Tissue samples

80
Q

Fixatives used in tissue sample for nucleic acid detection which are considered least damaging

A

10% buffered formalin
Acetone
Alcohol

81
Q

Cultures of bacteria, fungi, or virus may be analyzed for identification and characterization

A

Microbial cultures

82
Q

May still contain enough DNA or RNA to be detectable when amplification is employed

A

Cell free samples (ex: Serum)

83
Q

Mitochondrial DNA can be isolated from:

Nucleated/ Non-nucleated cells

A

Non-nucleated cells

84
Q

mRNA are compose of

A

Exons
Introns

85
Q

Contains instructions of making DNA/structural coding

A

Exons

86
Q

Nonstructural coding

A

Introns

87
Q

6 Types of Release/Lysis Methods

A

1) Mechanical method (grinding/mincing)
2) Heating/Boiling
3) Strong alkali (NaOH)
4) Sonication
5) Enzyme/Protease
6) Detergent (SDS, Triton X)

88
Q

Lysis method that use mortar and pestle

A

Mechanical method

89
Q

Lysis method that is most convenient and cheap method

A

Heating/Boiling

90
Q

Lysis method that use reagent kit and sodium hydroxide

A

Strong alkali (NaOH)

91
Q

Lysis method that se soundwaves

A

Sonication

92
Q

6 DNA Purification/Isolation method

A

Centrifugation
Organic Isolation
Inorganic Isolation
Solid-Phase Isolation
Chelating Ion Exchange Resin
Magnetic Beads

93
Q

Most efficient but toxic isolation method

A

Organic Isolation (Phenol/Chloroform Isoamyl Method)

94
Q

Primary reagent used in Organic Isolation that dissolves out lipids and proteins

A

Phenol
Chloroform

95
Q

In organic isolation, DNA will be found in the

A

Upper aqueous layer

96
Q

Use to precipitate out nucleic acid in organic isolation

A

Alcohol

97
Q

In organic isolation, after centrifuge,DNA/RNA will form a

A

pellet

98
Q

Uses low pH and high salt conditions to precipitate out proteins,leaving DNA in solution

A

Inorganic isolation(salting-out)

99
Q

In Inorganic isolation,DNA is later precipitated using

A

Alcohol

100
Q

Inorganic isolation is AKA

A

Salting-out

101
Q

Most common use DNA isolation

A

Solid-Phase Isolation

102
Q

Solid-Phase Isolation is based from?

A

Silica

103
Q

Silica-based matrices in the form of beads or colums are used to bind DNA

A

Solid-phase isolation

104
Q

To bind the nucleic acids to the silica matric, you can use

A

Chaotropic salts (guanine hydrochloride)

105
Q

Cause DNA to bind to silica by weakening H bind with water and forming cation bridges

A

Chaotropic salts

106
Q

In solid-phase isolation, DNA is eluted from the silica matric using a

A

Low salt buffer
Deionized water

107
Q

Major steps in Solid-Phase Isolation

A

Lysis
Binding
Washing
Elution

108
Q

A suspension of 5-10% chelex resin beads is mixed woth the specimen then boiled for lysis

A

Chelating Ion Exchange Resin (Chelex) Isolation

109
Q

How many percent of chelex resin beads suspension is needed

A

5-10%

110
Q

DNA are made to bind to magnetic beads coated with silicone dioxide

A

Magnetic Beads

111
Q

DNA are made to bind to magnetic beads coated with

A

silicone dioxide

112
Q

In magnetic bead, it is use to facilitate binding

A

Strong salts
Alcohol

113
Q

Magnetic particles are added to sample and bind to target molecule

Bind/ Wash/ Elute?

A

Bind

114
Q

Magnetic beads are captured and remainder of sample is washed away

Bind/ Wash/ Elute?

A

Wash

115
Q

Target molecule is released from magnetic particles

Bind/ Wash/ Elute?

A

Elute

116
Q
A

RMT (3 decks)

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