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A/ AH Biology: Unit 1 > Key Area 1: Laboratory Techniques for Biologists > Flashcards

Key Area 1: Laboratory Techniques for Biologists Flashcards

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1
Q

What are two things that can be intrinsically harmful in a lab?

A

Chemicals and organisms

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2
Q

What three things are at risk in a lab?

A
  • people
  • environment
  • other organisms
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3
Q

What are five control measures in order of most to least preferred?

A
  • elimination: replace the hazardous substance
  • substitution: use alternatives
  • isolation: contained environment
  • education: train people
  • personal protective equipment: last line of defence
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4
Q

What are risk assessments?

A

Carried out to ensure that working conditions are safe by identifying hazards and minimising risks through implementation of control procedures

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5
Q

What are devices for measuring liquids?

A
  • burette: titrations accurate for 1-100cm3
  • pipette: 30microlitres to 2cm3
  • syringe: 0.5microlitres to more than 50cm3
  • autopipette: 1microlitre to 20cm3
  • measuring cylinder: 5-2000cm3
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6
Q

What is a serial dilution?

A

Stepwise dilution where the dilution factor at each step is constant

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7
Q

What formula is used to calculate dilutions?

A

V1C1=V2C2

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8
Q

What is a linear dilution?

A

Even quantity separations

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9
Q

What is a log dilution?

A

Increase of factor ten each time

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10
Q

What are linear solutions used for?

A

Standard curves

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11
Q

What are long solutions used for?

A

Culturing microbes

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12
Q

What is a standard curve?

A

A graph that shows known concentrations and are used to determine unknown concentrations

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13
Q

HOw can standard curve concentrations be measured?

A

Colorimeter

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14
Q

How can pH be tested in an experiment?

A
  • universal indicator
  • pH paper
  • pH probe
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15
Q

How can you control pH?

A

With a buffer solution which resists a change in its pH when small amounts of acid or base is added into it

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16
Q

How does centrifugation seperate?

A

Density

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17
Q

What is the pellet?

A

The solid that forms at the bottom of the vessel in centrifugation

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18
Q

What is the supernatant?

A

The remaining liquid in centrifugation

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19
Q

What are three types of chromatography?

A

Thin layer
Affinity
Paper

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20
Q

What is thin layer chromatography?

A

Seperate by solubility on glass or plastic as amino acids with differing solubility travel different distances up the solid support

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21
Q

What is affinity chromatography?

A

Separates by specific bindingbetween molecules, separating biochemical mixtures based on a highly specific interaction such as that between antigen and antibody

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22
Q

What is paper chromatography?

A

Separates by solubility on paper, amino acids with different solubilitys travel different distances up the paper support being used

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23
Q

What is responsible for the separation of molecules in chromatography?

A

Solubility

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24
Q

How does electrophoresis separate?

A

By charge of size

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25
Q

How does electrophoresis work?

A

Proteins travel different distances through a gel with different charges at either end

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26
Q

What are two factors that effect the migration of proteins in a gel?

A

Charge and size

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27
Q

What is the separating factor in iso-electric focussing?

A

PH

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28
Q

How does iso-electric focussing work?

A

Protein no longer migrates at the pH where it has no overall charge and the protein precipitates out of solution

29
Q

Where does the protein precipitate out of solution and can be identified?

A

ISO-electric point

30
Q

What are antibody techniques used for?

A

Detecting and identifying proteins

31
Q

Why are antibodies suitable for detecting and identifying proteins?

A

They fight infection by specific binding to only one protein

32
Q

What are monoclonal antibodies?

A

Antibodies created from identical cells that attach to specific molecules

33
Q

What two cells fuse together to produce monoclonal antibodies?

A

Myeloma and B-lymphocytes

34
Q

What is the purpose of B-lymphocyte?

A

Produces the antibody

35
Q

What is the purpose of the myeloma?

A

Reproduce continuously without need of control

36
Q

What is the name given to the cell made by fusion of B-lymphocyte and myeloma?

A

Hybridoma

37
Q

What is used in the fusion process to produce the hybridoma?

A

Polyethylene glycol (PEG)

38
Q

What is immunoassay?

A

Based on antigen/antibody bonding principles and can be used to detect specific proteins

39
Q

What are the three types of Elisa?

A
  • direct Elisa
  • indirect Elisa
  • capture Elisa
40
Q

What is direct Elisa?

A

Antibody attaches directly to protein of interest

41
Q

What is indirect Elisa?

A

Secondary antibody with reporter enzyme attaches to secondary antibody that is attached to protein of interest

42
Q

What is capture Elisa?

A

Primary antibody added first, then protein that attaches to primary antibody, then secondary antibody that attaches to protein, then another antibody with a reporter enzyme attaches to the secondary antibody

43
Q

How do reporter enzymes detect specific proteins?

A

Observable colour changes

44
Q

How can antibodies be labelled for detection?

A

Reporter enzymes or fluorophore

45
Q

What is protein blotting?

A

After electrophoresis, the separates proteins are blotted onto a membrane which is treated with fluorescent malt labelled specific antibodies

46
Q

What is immunohistochemical staining?

A

Tissue is stained with specific labelled antibodies

47
Q

What is examined with brightfield microscopy?

A

Whole organisms, parts of organisms or thin sections of dissected tissue

48
Q

What is fluorescence microscopy?

A

Detecting specific proteins that have been bound to fluorescentkt labelled antibodies

49
Q

What is aeseptic technique?

A

Procedure used to ensure the environment is sterile

50
Q

What are five ways of maintaining a sterile environment?

A
  • sterilisation of equipment
  • disinfection if working area
  • wire loop flames before use
  • culture dish sealed
  • petri dish lid only partly opened at an angle
51
Q

What is the innoculum?

A

The original stock of cells used to innoculate the culture medium

52
Q

What is the explant?

A

The innoculum taken from plant or animal cell (check types notes)

53
Q

What is a haemocytometer used for?

A

Calculating cell counts

54
Q

What is trypan blue dye used for?

A

Staining cells

55
Q

What type of cells does trypan blue dye stain?

A

Only dead cells as the living cells can push out the dye

56
Q

What is vital stain?

A

Dye is only taken up by non-viable cells so viable cells are easily counted

57
Q

What is viable cell count?

A

Number of living cells in a sample

58
Q

What is non-viable cell count?

A

The number of dead cells in a sample

59
Q

What is total cells count?

A

Number of both living and dead cells in a sample

60
Q

What is a simple culture medium?

A

Allows conditions for gas exchange, a suitable pH and temperature and a suitable growing surface

61
Q

What is a complex growth medium?

A

Different nutrients are added to growth media depending on what you are growing

62
Q

What are the contents of a complex culture medium?

A
  • growth factor
  • serum
  • glucose/food sources
  • pH buffers
  • auxins
63
Q

What is the growth factor used for culturing plant cells?

A

Oxins

64
Q

Why is serum used in a complex culture medium?

A

It’s a source of growth factors

65
Q

What is proliferation?

A

The cell can divide

66
Q

What is a mono layer when culturing mammal cells?

A

A single layer of cells flat on the top of the surface

67
Q

What are primary cell lines?

A

Normal cells that only divide a certain number of times then die

68
Q

What are cancerous cell lines?

A

Can grow and divide infinitely without normal control of division or need for modification

A/ AH Biology: Unit 1 (6 decks)

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