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Advanced higher biology > 1.1 lab techniques > Flashcards

1.1 lab techniques Flashcards

1
Q

Hazard definition

A

Anything which can cause harm to a person

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2
Q

Hazards examples

A

Corrosive chemicals
Heat or flammable substances
pathogenic organisms
Mechanical equipment

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3
Q

COSSH

A

Control of substances hazardous to health

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3
Q

CLEAPSS

A

consortium of local educational authority for the provision of science services

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4
Q

Risk definition

A

The likelihood of a hazard occurring

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5
Q

What is in COSHH

A

chemicals
Nanotechnology
Biological agents

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6
Q

Standard PPE

A

Safety goggles
Lab coat
Gloves
Long trousers
Closed toe shoes

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7
Q

Linear dilutions

A

A series of solutions which differ in concentration by an equal interval

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8
Q

Log Dilutions

A

A series of solutions which differin concentration by a constant proportion

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9
Q

Making up linear dilutions

A

Add equal intervals of solution into each test tube and make up to the same volume with water

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9
Q

making up log dilutions

A

add 1cm of solution from sample into test tube and make up to 10cm3, then use 1cm3 of test tube 1 and make up to 10cm3 in test tube 2 and repeat

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9
Q

Semi log graphs

A

A graph with a linear axis and a log axis

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9
Q

Advantages of linear dilution

A

Each concentration is made up individually so errors only affect one solution.

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10
Q

Application of log dilution

A

Log dilution down the concentration of a bacteria solution
Then count her number of colonies on an agar plate.
Multiply up and you can find the concentration of the original solution

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11
Q

Buffer solution definition

A

A solution which remains at a constant pH when small quantities of acid or base are added

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12
Q

colourimetry uses

A

percentage transmission used to measure turbidity
absorbtion to measure colour and concentration

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13
Q

What is a centrifuge used for

A

Separation of substances of different density

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14
Q

more dense substance in centrifuge

A

will form a pellet at the bottom of the tube.

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15
Q

What happens to the less dense substance in a centrifuge

A

It remains in the supernatant

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16
Q

Supernatant

A

The solvent in a centrifuge

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16
Q

Types of chromatography

A

Paper
Thin layer
Affinity

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17
Q

Paper chromatography

A

Uses a special type of paper as the chromatogram, since paper contains cellulose which is polar it will allow for non polar molecules to travel further up the paper.

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18
Q

Thin layer chromatography

A

Uses silica gel, cellulose or an absorbent material.

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19
Q

Affinity chromatography

A

Used to separate one specific protein from a mixture by sending the mixture through a column with specific antibodies which will bind to the specific protein. This will mean the specific protein remains in the column whilst other proteins pass through.

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20
Q

Chromatography purpose

A

To be able to separate and purify the components of a mixture

21
Q

Chromatography basic principle

A

Draw a line on your chromatogram a the bottom and apply substance in a dried dot on the line
Place the bottom of the chromatogram in the solvent and put into a chromatography chamber, allow for the solvent front to travel up the chromatogram in the mobile phase.

22
Q

What affects the RF value of a substance in chromatography

A

The solubility of the substance in the solution and the ability of the substance to bind to the stationary phase

22
Q

Rf value

A

Distance traveled by the dot/distance traveled by solvent front

23
Q

gel electrophoresis

A

Where a protein is separated from a mixture/solution using electricity.

24
Q

Factors affecting SDS page electrophoresis

A

Based on size

24
Q

Factors affecting native gel electrophoresis

A

Size shape or charge

24
Q

Types of gel electrophoresis

A

Native gel and SDS page

25
Q

Native gel electrophoresis

A

Native gels analyse the proteins in their folded state without denaturing them

25
Q

SDS page electrophoresis

A

Gives each protein a negative charge and denatures them to separate the proteins out based on size.

26
Q

Separation by size alone rule

A

Small proteins travel further along the electrophoresis

27
Q

ELIZA definition

A

A technique which uses antibodies to bind to a specific antigen, which allows a reporter enzyme to produce a colour change and indicate the antigens presence.

27
Q

Isoelectric point definition

A

The pH that a protein’s overall charge becomes neutral and it falls out of solution

28
Q

Immunoassay

A

Where a specific antibody is used to bind to an antigen on a protein, which allows a reporter enzyme to produce a colour change. This is done to identify a specific protein in a mixture.

29
Q

ELISA stands for

A

Enzyme linked immunosorbent assay

30
Q

Direct ELISA

A

antigen binds to a multi well plate
Antibody binds to the antigen
A reporter enzyme is added to then well and a colour change occurs.

30
Q

Types of ELISA

A

direct ELISA
Indirect ELISA
sandwich ELISA

31
Q

Indirect ELISA

A

An antigen will bind to a multi well plate
The primary antibody will bind to the antigen
The secondary antibody binds to the primary antibody
The reporter enzyme is added and causes a colour change

32
Q

Sandwich ELISA

A

A primary antibody binds to a multi well plate
An antigen binds to the primary antibody
A secondary antibody binds to the antibody
The reporter enzyme linked to the second antibody causes colour change.

33
Q

When is western blotting used

A

After SDS page electrophoresis

34
Q

What is western blotting

A

Where after an electrophoresis proteins are transferred to a solid medium

34
Q

Protein blotting

A

Western blotting using proteins

35
Q

Fluorescent microscopy

A

Where fluorescent light emitting chemicals are used to bind to proteins so they can be identified under a microscope.

35
Q

Bright field microscopy

A

A where a sample is put under a microscope and a white light is shined through the sample

35
Q

Bright light microscopy samples

A

Whole organisms, parts of organisms, sections of tissues and individual cells

35
Q

Immunofluorescence

A

Where antibodies are used to fluorescent tag a protein.

36
Q

Cell culture

A

Growing cells in a lab

37
Q

Culture media contents

A

Water
Salts
Amino acids
Vitamins
Glucose

38
Q

Animal cell culture requirements

A

Growth factors

39
Q

Aseptic techniques

A

Eliminate and kill all unwanted microorganism contamination when culturing microorganisms

40
Q

Precautions taken when working with cultures

A

Sterilise work area with cleaning products before
Good personal hygiene at all times
Sterile reagents and media must be used
Sterile handling must occur such as wiping down containers with ethanol, flaming containers when opened and exposing the all reagents to air for as little time as possible.

41
Q

Haemocytometer

A

A piece of apparatus which create a 0.1 mm chamber with a grid on the side.

42
Q

Total cell count

A

Count both viable and dead cells

42
Q

Dye for dead cells

A

Trypan blue

42
Q

Live cell count

A

Stain dead cells with trypan blue then count the colourless cells.

43
Q

Percentage viability

A

Live cells/ total cells x100

Advanced higher biology (11 decks)

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