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AH Biology > 1.1 Laboratory Techniques for Biologists > Flashcards

1.1 Laboratory Techniques for Biologists Flashcards

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1
Q

Hazard

A

Something that can cause harm

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2
Q

Something that can cause harm

A

Hazard

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3
Q

What can hazards arise from

A

Toxic chemicals and corrosive chemicals

Heat and flammable substances

Pathogenic organisms

Mechanical equipment

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4
Q

Toxic chemicals and corrosive chemicals

Heat and flammable substances

Pathogenic organisms

Mechanical equipment

A

What hazards arise from

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5
Q

Risk

A

The likelihood of harm arising from exposure to any hazard

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6
Q

The likelihood of harm arising from exposure to any hazard

A

Risk

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7
Q

Risk assessment

A

Identify and evaluate the hazards

AND

identify controls to minimise risks

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8
Q

Identify and evaluate the hazards

AND

identify controls to minimise risks

A

Risk assessment

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9
Q

Control measures to limit risks

A

Wear PPE
- safety goggles, lab coat, no open toe shoes

Appropriate handling techniques

Use aseptic techniques
- to minimise microbial contamination

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10
Q

Wear PPE
- safety goggles, lab coat, no open toe shoes

Appropriate handling techniques

Use aseptic techniques
- to minimise microbial contamination

A

Control measures to limit risk

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11
Q

How to ensure accuracy of experiments and data generated

A

Using the most appropriate apparatus and equipment

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12
Q

Using the most appropriate apparatus and equipment ensures what in practical experiments

A

Accuracy of experiments and data generated

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13
Q

What must be done to measure volumes accurately

A

Select the appropriate apparatus

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14
Q

When is linear dilution used

A

When the substance being diluted is the independent variable in the experiment

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15
Q

Linear dilutions vary from eachother in _________

A

Equal interval

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16
Q

______ dilutions differ from eachother in equal interval

A

Linear

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17
Q

How to make a linear dilution

A

start with a stock solution of a known concentration

add increasing volume of that solution to separate test tubes

then a pure solvent (eg, distilled water) so that an equal volume of each dilution is produced

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18
Q

What can be used to produce a standard curve

A

A linear dilution series

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19
Q

What must you do to the instrument when making a standard curve from linear dilution

A

The instrument must be calibrated using a blank as a baseline

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20
Q

What can standard curves be used for

A

To determine the unknown concentration of a solution

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21
Q

What can be used to determine the unknown concentration of a solution

A

Standard curve

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22
Q

What is often used in microbiology to estimate the concentration or density of cells in a stock culture

A

Log dilution

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23
Q

What is log dilution often used for

A

In microbiology to estimate the concentration or density of cells in a stock culture

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24
Q

Log dilutions differ by a _____________

A

Constant proportion

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25
Q

_____ dilutions differ by a constant proportion

A

Log

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26
Q

How are log dilutions made

A

Created by diluting a stock solution by a factor

then further diluting the dilution produced by the same factor, keeping the pure solvent constant.

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27
Q

What does colorimetry measure

A

The the concentration of pigments in a solution

Turbidity

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28
Q

Buffer

A

Solutions that can resist changes of pH, even though acid or alkali is added

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29
Q

Solutions that can resist changes of pH, even though acid or alkali is added

A

Buffer

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30
Q

What do buffers allow for

A

pH of a mixture to be kept constant to prevent enzymes from denaturing and affecting its structure which reduces affinity for the substrate

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31
Q

What keeps the pH of a mixture to be kept constant to prevent enzymes from denaturing and affecting its structure which reduces affinity for the substrate

A

Buffer

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32
Q

Separation techniques for solubility

A

Filtration

Chromatography
- paper
- thin layer
- affinity

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33
Q

Filtration

Chromatography
- paper
- thin layer
- affinity

A

Separation techniques for solubility

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34
Q

Separation techniques for size

A

Centrifugation

Gel electrophoresis

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35
Q

Centrifugation

Gel electrophoresis

A

Separation techniques for size

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36
Q

Separation techniques for charge

A

Isoelectric point

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37
Q

Isoelectric point

A

Separation techniques based on charge

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38
Q

Centrifugation separates based on what

A

Components of a suspension that have a different density

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39
Q

What separates components of a suspension that have a different density

A

Centrifugation

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40
Q

Separation of substances in centrifugation

A

Denser substances found in pellet at bottom

Less dense substances remain in the liquid, the supernatant

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41
Q

Paper chromatography technique

A

Add a spot of solution to origin line of the chromatography strip

Dip in appropriate solvent and allow to run

Develop it with an amino acid stain

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42
Q

Add a spot of solution to origin line of the chromatography strip

Dip in appropriate solvent and allow to run

Develop it with an amino acid stain

A

Paper chromatography method

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43
Q

Stationary phase in paper chromatography

A

Paper

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44
Q

Paper in paper chromatography

A

Stationary phase

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45
Q

Mobile phase in paper chromatography

A

Water

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46
Q

Water in paper chromatography

A

Mobile phase

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47
Q

Rf value

A

A measure of the distance travelled by a component/ water travelled by the solvent

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48
Q

A measure of the distance travelled by a component/ water travelled by the solvent

A

Rf value

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49
Q

Larger Rf value means what

A

Component is more soluable

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50
Q

Smaller Rf value

A

Component is less soluable

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51
Q

What does the speed that each solute travels along the chromatogram depend on

A

It’s differing solubility in the solvent used

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52
Q

what depends on the differing solubility in the solvent used

A

The speed that each solute travels along the chromatogram

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53
Q

What can chromatography be used to identify samples of

A

Drugs and lipstick

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54
Q

Thin layer chromatography method

A

A glass plate with a thin layer of silica gel is used

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55
Q

A glass plate with a thin layer of silica gel is used

A

Thin layer chromatography

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56
Q

Soluble gel in thin layer chromatography

A

Stationary phase

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57
Q

Stationary phase in thin layer chromatography

A

Soluble gel

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58
Q

Mobile phase in thin layer chromatography

A

Solvent

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59
Q

Solvent in thin layer chromatography

A

Mobile phase

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60
Q

What does ninhydrin do

A

Turns colourless samples purple

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61
Q

In affinity chromatography, what does separation occur between

A

Specific interaction between 2 molecules

62
Q

Affinity chromatography

A

A solid matrix or gel column is created with specific molecules bound to the matrix or gel

Soluble, target proteins in a mixture, with a high affinity for these molecules, become attached to them as the mixture passes down the column

Other non-target molecules with a weaker affinity are washed out.

63
Q

A solid matrix or gel column is created with specific molecules bound to the matrix or gel

Soluble, target proteins in a mixture, with a high affinity for these molecules, become attached to them as the mixture passes down the column

Other non-target molecules with a weaker affinity are washed out.

A

Affinity chromatography

64
Q

Buffers in affinity chromatography

A

Buffers with different pH removes molecules of interest or competitive inhibitors are used, then removed by dialysis

65
Q

Gel electrophoresis

A

Charged macromolecules move through an electric field applied to a gel matrix

66
Q

A process used to separate proteins and nucleic acids

A

Gel electrophoresis

67
Q

Gel electrophoresis method

A

Charged macromolecules move through an electric field applied to a gel matrix

68
Q

Charged macromolecules move through an electric field applied to a buffer gel matrix towards a positively charged electrode

A

Gel electrophoresis

69
Q

Native PAGE gels in gel electrophoresis

A

do not denature the molecules being separated

they are separated by shape, size and charge.

→tricky to carry out and structure and function is preserved

70
Q

do not denature the molecules being separated

they are separated by shape, size and charge.

→tricky to carry out and structure and function is preserved

A

Native PAGE gels

71
Q

SDS PAFE gels in gel electrophoresis

A

separate the proteins by size alone

They do this by giving all molecules an equally negative charge and denaturing them

→simple to carry out but structure and function of any protein may be lost

72
Q

separate the proteins by size alone

They do this by giving all molecules an equally negative charge and denaturing them

→simple to carry out but structure and function of any protein may be lost

A

SDS PAGE gels

73
Q

Isoelectric point

A

The pH value at which a surface protein is electrically neutral and will precipitate out of a solution

74
Q

The pH value at which a surface protein is electrically neutral and will precipitate out of a solution

A

Isoelectric point

75
Q

What can IEP be used for

A

To separate proteins from a mixture

76
Q

IEP method

A

If the solution is buffered to a specific pH

only the protein(s) that have an IEP of that pH will precipitate

77
Q

IEP method for separating a mixture of proteins

A

By a type of electrophoresis

Run the mixture through a pH gradient gel and each protein will precipitate at its isoelectric point.

78
Q

By a type of electrophoresis

Run the mixture through a pH gradient gel and each protein will precipitate at its isoelectric point.

A

IEP separating multiple proteins in a mixture

79
Q

Monoclonal antibodies

A

monospecific antibodies that are identical to each other because they are made by one particular group of B lymphocytes and bind to the same antigen

80
Q

monospecific antibodies that are identical to each other because they are made by one particular group of B lymphocytes and bind to the same antigen

A

Monoclonal antibodies

81
Q

Polyclonal antibodies

A

not identical to each other and are made from several B lymphocytes

82
Q

not identical to each other and are made from several B lymphocytes

A

Polyclonal antibodies

83
Q

What are antibodies used in

A

The immune system and are responsible for the identification of foreign particles or antigens and flagging them for destruction

84
Q

The immune system and are responsible for the identification of foreign particles or antigens and flagging them for destruction

A

Antibodies

85
Q

Immunoassay techniques

A

Used to detect and identify specific proteins

86
Q

Used to detect and identify specific proteins

A

Immunoassay techniques

87
Q

Immunoassay techniques principle

A

They use stocks of monoclonal antibodies, that bind to a specific antigen on the protein of interest and is linked to a chemical ‘label’

88
Q

They use stocks of monoclonal antibodies, that bind to a specific antigen on the protein of interest and is linked to a chemical ‘label’

A

Immunoassay techniques

89
Q

Chemical label

A

Reporter enzyme producing a colour change/chemiluminescence/fluorescence

90
Q

Reporter enzyme producing a colour change/chemiluminescence/fluorescence

A

Chemical label

91
Q

Techniques to detect proteins using antibodies

A

ELISA

western blotting

92
Q

Techniques to detect proteins using antibodies

A

ELISA

western blotting

93
Q

ELISA

western blotting

A

Techniques to detect proteins using antibodies

94
Q

ELISA

A

System used to detect specific antigens or antibodies

Can exist in two forms:

One that detects the presence of a particular antibody

The other detects the presence of a specific antigen

95
Q

ELISA

A

System used to detect specific antigens or antibodies

Can exist in two forms:

One that detects the presence of a particular antibody

The other detects the presence of a specific antigen

96
Q

System used to detect specific antigens or antibodies

Can exist in two forms:

One that detects the presence of a particular antibody

The other detects the presence of a specific antigen

A

Enzyme linked immunosorbent assay

97
Q

Uses for ELISA

A

HIV detection

98
Q

What is used for HIV detection

A

ELISA

99
Q

What is used after SDS PAGE

A

Western blotting

100
Q

When is western blotting used

A

After SDS PAGE

101
Q

Western blotting technique

A

The separated proteins from gel electrophoresis are transferred onto a solid membrane and dried

The proteins can be labelled by soaking the blot with specific antibodies, which can bind to specific proteins.

A second antibody with reporter enzymes are then added.

The presence of a protein can be visible to the eye following the addition of the reporter enzyme’s substance

102
Q

The proteins can be labelled by soaking the blot with specific antibodies, which can bind to specific proteins.

A second antibody with reporter enzymes are then added.

The presence of a protein can be visible to the eye following the addition of the reporter enzyme’s substance

A

Western blotting method

103
Q

Can you see the presence of a protein in western blotting

A

It is visible to the eye after the addition of the reporter enzyme’s substrate

The reporter will change colour when the antigen is present

104
Q

When might a false positive occur in western blotting

A

If a secondary antibody in still present in the well (due to insufficient washing)

This, the secondary antibodies can react with the substrate to produce the coloured product, despite not being bound to the primary antibody

105
Q

Bright field microscopy method

A

Light is transmitted through object and magnified by objective lens

Then by eyepiece lens before being observed

106
Q

Light is transmitted through object and magnified by objective lens

Then by eyepiece lens before being observed

A

Bright field microscopy method

107
Q

What can be observed with bright field microscopy

A

Whole organisms

Parts of organisms

Thin sections of dissected tissue

Individual cells

108
Q

What can be used to observe..

Whole organisms

Parts of organisms

Thin sections of dissected tissue

Individual cells

A

Bright field microscopy

109
Q

What does fluorescence microscopy observe

A

Particular protein structures

110
Q

What can be used to visualise particular protein structures

A

Fluorescence microscopy

111
Q

What can be used to visualise particular protein structures

A

Fluorescence microscopy

112
Q

Fluorescence microscopy method

A

Fluorescence molecules absorbs light of one colour and emits light of a different colour

113
Q

Fluorescence microscopy method

A

Fluorescence molecules absorbs light of one colour and emits light of a different colour

Protein structures have fluorescent tags added

114
Q

Fluorescence molecules absorbs light of one committee and emits light of a different colour

Protein structures have fluorescent tags added

A

Fluorescence microscopy

115
Q

What does aseptic techniques do

A

Eliminates unwanted microbial contaminants and prevents competition with target microbe

116
Q

What eliminates unwanted microbial contaminants and prevents competition with target microbe

A

Aseptic techniques

117
Q

What does aseptic technique involve

A

The sterilisation of equipment and culture media by heat or chemical means

And

Subsequent exclusion of microbial contaminants

118
Q

what involves the sterilisation of equipment and culture media by heat or chemical means

And

Subsequent exclusion of microbial contaminants

A

Aseptic techniques

119
Q

What does aseptic technique ensure

A

Cultures are pure

120
Q

What ensures cultures are pure

A

Aseptic techniques

121
Q

Microbial culture

A

A method of multiplying organisms by letting them reproduce in a culture medium under controlled lab conditions

122
Q

A method of multiplying organisms by letting them reproduce in a culture medium under controlled lab conditions

A

Microbial culture

123
Q

What can a microbial culture be started using

A

An inoculum of microbial cells on an agar medium, or a broth with suitable nutrients

124
Q

What can be started with an inoculum of microbial cells on an agar medium, or a broth with suitable nutrients

A

Microbial culture

125
Q

Specific culture media

A

Culture media that promote the growth of specific types of cells and microbes

126
Q

Culture media that promote the growth of specific types of cells and microbes

A

Specific culture media

127
Q

Animal cells are growth in what

A

Medium containing proteins

128
Q

What are grown in medium containing proteins

A

Animal cells

129
Q

What is the name for proteins in animal cell medium

A

Growth factors

130
Q

Where do growth factors come from

A

grSerum

131
Q

What do growth factors do

A

Promote cell growth and proliferation

Essential for the culture of most animal cells

132
Q

What promote cell growth and proliferation

Essential for the culture of most animal cells

A

growth factors

133
Q

How many times can primary lines divide by

A

A limited number of times

134
Q

What cell lines can divide a limited number of times

A

Primary cell lines

135
Q

What types of cells can divide an unlimited number of times

A

Tumour cell lines

136
Q

How many times can tumour cell lines divide

A

Unlimited divisions

137
Q

What is the benefit of plating out a liquid microbial culture onto a solid media

A

It allows the number of colony forming units to be counted

Thus the density of cells in culture can be estimated

138
Q

How could you count the number of colony forming units in a liquid microbial culture

A

Plating out the liquid onto solid media

139
Q

What is often needed to plate a liquid microbial culture onto a solid media

A

Serial dilution

140
Q

What can serial dilution be needed for

A

To achieve a suitable colony count

141
Q

Culture requirements

A

Aseptic conditions, incl. antibiotics

Solid surface/tissue culture flask (except blood cells)

Nutrients, eg. Glucose, amino acids, etc

Suitable gas exchange

Growth factors, generally are specific to type of cell

142
Q

Aseptic conditions, incl. antibiotics

Solid surface/tissue culture flask (except blood cells)

Nutrients, eg. Glucose, amino acids, etc

Suitable gas exchange

Growth factors, generally are specific to type of cell

A

Culture requirements

143
Q

Haemocytometers

A

Specialised microscope cells which are used to estimate cell numbers in a liquid culture

144
Q

Specialised microscope cells which are used to estimate cell numbers in a liquid culture

A

Haemocytometers

145
Q

What is needed to identify and count living viable cells

A

Vital staining

146
Q

What is viral staining used for

A

To identify and count living viable cells

147
Q

Trypan blue

A

Used to stain dead cells

148
Q

Used to stain dead cells

A

Trypan blue

149
Q

Disadvantages of Haemocytometers

A

dead cells are not distinguished form live cells (unless stained)

small cells can be difficult to locate

numbers obtained are only an estimate

time consuming

Clumping of cells, hard to count individually (leads to inaccuracy)

150
Q
A

AH Biology (9 decks)

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