lab techniques Flashcards

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1
Q

define risk

A

the likelihood of harm arising from exposure to a hazard

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2
Q

what are the types of hazard?

A

toxic or corrosive chemicals, heat or flammable substance, pathogenic organisms and mechanical equipment

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3
Q

what control measures are used to control risk

A

appropriate handling techniques,protective clothing and equipment and aseptic techniques

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4
Q

What does risk assessment involve?

A

involves identifying control measures to minimize the risk.

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5
Q

which dilution series differs by an equal interval

A

linear dilutions

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6
Q

which dilution series differs by a constant proportion

A

logarithmic dilutions

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7
Q

what is used to determine the concentration of a colored solution

A

Absorbance using suitable wavelength filters

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8
Q

what is used to determine turbidity

A

percentage transmission

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9
Q

what is a buffer solution

A

it is a mixture of compounds (often a weak acid and its base);

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10
Q

what is pH

A

it is a continuous scale and indicates the concentration of hydrogen ions

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11
Q

what is the purpose of a buffer

A

it resists small changes in pH, by the addition of an acid or alkalai

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12
Q

what does a colorimeter do

A

measures the passage of light through a solution or the absorbance of light by the solution

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13
Q

what is the affect of the addition of an acid or alkali on a buffer

A

Addition of acid or alkali has very small effects on the pH of a buffer, allowing the pH
of a reaction mixture to be kept constant.

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14
Q

what is a standard curve used for

A

Plotting measured values for known concentrations to produce a line or curve
allows the concentration of an unknown to be determined from the standard curve.

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15
Q

Method and uses of a colorimeter to quantify
concentration and turbidity

A

Calibration with appropriate blank as a baseline; use of absorbance to determine
concentration of a coloured solution using suitable wavelength filters; use of percentage transmission to determine turbidity, such as
cells in suspension.

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16
Q

how does a centrifuge separate components in a mixture

A

More dense components settle in the pellet; less dense components remain in the
supernatant.

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17
Q

what does the speed that each solute travel along the chromatogram depend on

A

The speed that each solute travels along the
chromatogram depends on its differing
solubility in the solvent used.

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18
Q

what is paper thin chromatography used for

A

Paper and thin layer chromatography can be
used for separating different substances such
as amino acids and sugars

19
Q

what is a centrifuge used for

A

centrifuge to separate substances of
differing density

20
Q

Principle of affinity chromatography and its
use in separating proteins

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.

21
Q

Principle of gel electrophoresis

A

Charged macromolecules move through an
electric field applied to a gel matrix. it is used to seperate protiens and nucleic acids

22
Q

how do native gels seperate molecules

A

Native gels do not denature the molecule so
that separation is by shape, size and charge.

23
Q

SDS–PAGE separates proteins

A

SDS–PAGE gives all the molecules an
equally negative charge and denatures them,
separating proteins by size alone.

24
Q

define iso-electric point

A

IEP is the pH at which a soluble protein has
no net charge and will precipitate out of
solution.

25
Q

If the solution is buffered to a specific pH, only the what will precipitate out

A

If the solution is buffered to a specific pH, only the protein(s) that have an IEP of that pH will precipitate

26
Q

Soluble proteins can be separated using an
electric field and a what

A

Soluble proteins can be separated using an
electric field and a pH gradient. A protein
stops migrating through the gel at its IEP in the pH gradient because it has no net charge.

27
Q

what are immunoassay techniques used to detect

A

Immunoassay techniques are used to detect
and identify specific proteins.These techniques use stocks of antibodies
with the same specificity, known as
monoclonal antibodies

28
Q

An antibody specific to the protein antigen is linked to a chemical ‘label’. what is this ‘label’

A

An antibody specific to the protein antigen is linked to a chemical ‘label’. The ‘label’ is often a reporter enzyme producing a colour change, but chemiluminescence, fluorescence and other
reporters can be used.

29
Q

in some cases assay uses what

A

In some cases the assay uses a specific
antigen to detect the presence of antibodies.

30
Q

what is western blotting

A

Western blotting is a technique, used after
SDS–PAGE electrophoresis
The separated proteins from the gel are
transferred (blotted) onto a solid medium.

31
Q

how can proteins be identified

A

The proteins can be identified using specific antibodies that have reporter enzymes
attached

32
Q

what is bright feild commonly microscopy used for

A

Bright-field microscopy is commonly used to
observe whole organisms, parts of organisms, thin sections of dissected tissue
or individual cells

33
Q

what is flourescence microscopy

A

Fluorescence microscopy uses specific
fluorescent labels to bind to and visualise
certain molecules or structures within cells or
tissues

34
Q

what do aseptic techniques do

A

Aseptic technique eliminates unwanted
microbial contaminants when culturing microorganisms or cells

35
Q

what do aseptic techniques involve

A

Aseptic technique involves the sterilisation of
equipment and culture media by heat or
chemical means and subsequent exclusion of
microbial contaminants.

36
Q

how can a microbial culture be started

A

A microbial culture can be started using an
inoculum of microbial cells on an agar
medium, or in a broth with suitable nutrients

37
Q

many culture media that exist do what

A

Many culture media exist that promote the
growth of specific types of cells and
microbes.

38
Q

Animal cells are grown in medium containing what

A

Animal cells are grown in medium containing
growth factors from serum

39
Q

what are growth factors

A

Growth factors are proteins that promote cell
growth and proliferation. Growth factors are
essential for the culture of most animal cells.

40
Q

In culture, primary cell lines can divide a
limited number of times, whereas what

A

In culture, primary cell lines can divide a
limited number of times, whereas tumour
cells lines can perform unlimited divisions

41
Q

Plating out of a liquid microbial culture on
solid media allows what

A

Plating out of a liquid microbial culture on
solid media allows the number of colony forming units to be counted and the density of cells in the culture estimated

42
Q

what is serial dilution often needed for

A

Serial dilution is often needed to achieve a
suitable colony count

43
Q

what is a haemocytometer used for

A

to estimate cell numbers in a liquid culture

44
Q

what is vital staining required for

A

Vital staining is required to identify and count
viable cells