Unit 1 - Lab techniques
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Describe how to carry out a serial dilution | 1ml of stock solution with 9ml of water. Then 1ml of diluted solution with 9ml of water and so on.
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What substance controls pH | buffers
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Describe two techniques that can be used to quantify the concentration of an unknown solution | Colorimetry and Titration
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What is a standard curve | Measuring the absorbance of known concentrations in a colorimeter to extrapolate unknown from a graph
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Which property allows substances to be separated in centrifugation | density
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What name is given to the substance at the bottom and top during centrifugation | bottom - pellet
top - supernatant
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small substances are found where after centrifugation | in the supernatant
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What techniques separate proteins based on pH | Iso electric focussing
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What is meant by the iso electric point | When proteins have an OVERALL neutral charge and precipitates out of solution
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What are the two types of gel electrophoresis by which proteins can be separated | Denaturing and native/non denaturing
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Explain the process of denaturing gel electrophoresis | All proteins are denatured to give a negative charge and move to positive end dependent on size
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Explain the process of non denaturing gel electrophoresis | Proteins remain in original conformation (no denaturing) and migrate through gel dependent on SIZE & CHARGE
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What type of gel electrophoresis depends only on size | Denaturing (as all proteins are artificially made negative)
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What type of gel electrophoresis depends on size and charge | Native/non denaturing (protein charge not affected)
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Name a factor affecting protein migration apart from size or charge | size of pores in buffer
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What are the types of microscopy | 1. bright field 2. Fluorescence
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Which type of microscopy allows tiny structures such as individual proteins within the cell to be viewed | fluorescence
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Which type of microscopy is more limited in its resolution | bright field
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Which type of microscopy is more simple to use | bright field
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Which type of microscopy involves antibodies to tag objects | fluorescence
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Name an object that can be viewed under a bright field microscope | whole organisms, parts of organism or thin tissues
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What does a haemocytometer estimate | total cell count
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What technique is able to distinguish between dead cells and viable cells | Vital Staining with trypan blue
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Explain why only dead cells stain with trypan blue | Trypan blue toxic so actively pumped out by living (viable) cells
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Explain how to estimate total cell count in a haemocytometer | Ensure all units are in CM. L x B X Depth = volume Count cells that fit in grid Proportion calculation to scale up for chosen volume which is always larger than volume calculated
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What term is given to cells that can only divide a limited number of times (hayflick) limit | primary cell lines
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What term is given to cells that can divide infinitely | cancer cells
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Why are growth regulators/hormones used in plant tissue | To enable explant to divide into callus AND to enable callus to differentiate into plantlet
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Why do mammalian cells need foetal bovine serum | For cell proliferation
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Why do mammalian cells need antibiotics | to kill bacteria which grow more readily than mammalian cells
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What type of organisms are produced from inoculum | microbes
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What type of organisms are produced from explants | plants
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Name a plant hormone/growth regulator | auxin or cytokinins
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What two cells fuse to make a hybridomas | B lymphocyte with a myeloma cell
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How are specific monoclonal antibodies produced in a lab | injecting mouse with specific antigen that produces antibodies. Removing spleen and extract B lymphocytes & fusing with myeloma cell to form hybridomas
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What substance are the hybridomas placed in and subsequently diluted to ensure one cell per well | polyethylene glycol (PEG).
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