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Advanced Higher Biology > 1.1 Laboratory Techniques for Biologists > Flashcards

1.1 Laboratory Techniques for Biologists Flashcards

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1
Q

What are the three things that can cause a hazard in a laboratory?

A

Substances, organisms and equipment

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2
Q

Hazards in the lab can include:

A

toxic chemicals, heat and flammable substances, pathogenic organisms, and mechanical equipment

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3
Q

What is Risk?

A

Risk is the likelihood of harm arising from exposure to a hazard

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4
Q

What is a risk assessment?

A

A risk assessment involves identifying possible risks and the control measures to minimise them

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5
Q

What are control measures?

A

Control measures used to minimise risk include using appropriate handling technique.

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6
Q

How do the dilutions in a linear dilution series work?

A

Dilutions differ by an equal interval, for example 0.1m, 0.2m, 0.3m and so on

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7
Q

How do the dilutions in a log dilution series work?

A

Dilutions differ by constant proportion, for example 10^-1, 10^-2, 10^-3 and so on

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8
Q

How is a standard curve made and how is it used?

A

A standard curve is produced by plotting measured values for known concentrations; it is used to determine the concentration of an unknown solution

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9
Q

What are buffers?

A

Buffers are used to control pH; the addiction of an acid or alkali only has a very small effect on its pH, allowing the pH of a reaction mixture to be kept constant

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10
Q

What is a colorimeter used for?

A

A colorimeter can be used to quantify the concentration and turbidity of a solution.

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11
Q

How can a colorimeter be calibrated?

A

The colorimeter is calibrated using an appropriate blank as a baseline, for example; deionised water

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12
Q

What do colorimeters measure?

A

They measure absorbance to determine the concentration of a coloured solution using suitable wavelength filters. The measurement of percentage transmission is used to determine turbidity

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13
Q

What is centrifugation?

A

Centrifugation is a technique used to separate substances of differing density.

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14
Q

Where do more dense components settle?

A

They settle in the pellet

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15
Q

Where do less dense components remain?

A

They remain in the supernatant

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16
Q

What is Paper and Thin Layer Chromatography (TLC) used for?

A

They are used for separating different substances such as amino acids and sugars.

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17
Q

How does Paper and Thin Layer Chromatography work?

A

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

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18
Q

What is Affinity Chromatography used for?

A

It’s a separation technique in which soluble target proteins with a high affinity in a mixture become attached to specific molecules as the mixture passes down a column. Non-target molecules with a weaker affinity are washed out.

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19
Q

What is Gel Electrophoresis?

A

It is a technique that can be used to separate proteins and nucleic acids.

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20
Q

How does Gel Electrophoresis work?

A

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

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21
Q

What is Native Gel Electrophoresis?

A

Separates proteins and nucleic acid by their shape, size and charge.

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22
Q

What does Native Gel Electrophoresis do that normal Gel Electrophoresis doesn’t do which allows it to maintain shape, size and charge?

A

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

23
Q

What is SDS-PAGE and what is the difference between it and Native and normal gel electro?

A

SDS-PAGE gives all the molecules an equally negative charge and denatures them, separating proteins by size alone. Which is an easier technique to conduct than native gel electrophoresis

24
Q

Proteins can be separated from a mixture using what?

A

Their isoelectric points (IEPs)

25
Q

What is an isoelectric point?

A

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

26
Q

What happens if the solution is buffered to a specific pH?

A

Only the proteins that have an IEP at that pH will precipitate.

27
Q

Soluble proteins can be separated using what?

A

An electric field and a pH gradient gel.

28
Q

Why does a protein stop migrating through the gel at it’s IEP?

A

As at it’s IEP in the pH gradient it has no net charge.

29
Q

What can be used to detect proteins?

A

Proteins can be detected using antibodies.

30
Q

What are Immunoassay techniques used for?

A

To detect and identify specific proteins.

31
Q

Immunoassay techniques use stocks of what?

A

Stocks of antibodies with the same specificity, known as monoclonal antibodies.

32
Q

An antibody specific to the antigen is linked to what?

A

A chemical ‘label’

33
Q

The label is often a ____ ____ producing a _____ ______, but what other things can be used?

A

The label is often a reporter enzyme producing a colour change, but chemiluminescence, fluorescence radioactivity and other reporters can be used?

34
Q

In some cases, the assay used what to detect the presence of antibodies?

A

A specific antigen

35
Q

What is Western Blotting?

A

Western blotting is technique used after SDS-PAGE electrophoresis. In western blotting, the separated proteins from the gel are transferred (blotted) on to a solid medium.

36
Q

The proteins used in SDS-PAGE and Western blotting can be identified through?

A

Specific antibodies that have reporter enzymes attached.

37
Q

Why does washing occur in immunoassays, and the implication for the results if not done properly?

A

To ensure all antigen that isn’t bind to an antibody is removed from the assay, if no antigen’s are bonded to antibodies and the antigen is still present the test will show a false positive.

38
Q

What does an immunoassay rely on?

A

The specificity of antibodies i.e. their ability to recognise and bind with only one antigen

39
Q

What is an example of an immunoassay technique?

A

ELIZA

40
Q

What is an ELISA (enzyme-linked immune absorbent assay) test?

A

An analytical technique which uses antibodies to detect the presence of an antigen within a solution

41
Q

What is bright-field microscopy commonly used for?

A

Observing whole organisms, parts of organisms, thin sections of dissected tissue of individual cells.

42
Q

What does fluorescence microscopy use fluorescent label for?

A

Fluorescent labels bind to and visualise certain molecules or structures within cells or tissues.

43
Q

Aseptic technique eliminates what?

A

Unwanted microbial contaminants when culturing micro-organisms in cells.

44
Q

Aseptic technique includes:

A

The sterilisation of equipment and culture media by heat or chemical means, and subsequent exclusion of microbial contaminants.

45
Q

An inoculum of microbial cells on an agar medium, or in a broth with suitable nutrients can start what?

A

A microbial culture

46
Q

Many culture media exist that promote what?

A

The growth of specific types of cells and microbes.

47
Q

Animal cells are grown in what?

A

A medium containing growth factors from serum.

48
Q

What are growth factors?

A

Growth factor are proteins that promote cell growth and proliferation.

49
Q

Growth factors are essential for the culture of most ____ ____.

A

Animal Cells

50
Q

State the difference between primary cell lines and tumour cells when diving

A

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

51
Q

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

A

The number of colony-forming units to be counted and the density of cells in the culture estimates.

52
Q

Serial dilution - whether it be linear or log dilution - is often needed to achieve what?

A

A suitable colony count.

53
Q

What are haemocytometers used for?

A

To estimate cell numbers in a liquid culture

54
Q

Vital Staining is used for?

A

To identify and count viable cells

Advanced Higher Biology (5 decks)

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