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Advanced Higher Bio

Unit 1 lab techniques

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....
What substance controls pH buffers
Describe two techniques that can be used to quantify the concentration of an unknown solution 1.colorimetry 2. titration
What is another term for a colorimeter spectrophotometer
What is a standard curve measuring absorbance of known concentrations in a colorimeter to extrapolate unknown from graph
On what property are substances separated in centrifugation density
What name is given to the substance at bottom and top during centrifugation bottom - pellet top - supernatant
Small substances are found where after centrifugation in supernatant
What technique separates proteins based on pH Iso electric focusing
What is meant by the iso electric point of a protein When it is OVERALL neutral and precipitates out of solution
What are the two types of gel electrophoresis by which proteins can be separated 1. Denaturing 2. Native/non denaturing
Explain the process of denaturing gel electrophoresis All proteins are denatured to give a negative charge and move to positive end dependent on size
Explain the process of native gel electrophoresis Proteins remain in original conformation (no denaturing) and migrate through gel dependent on SIZE & CHARGE
What type of gel electrophoresis depends only on size Denaturing (as all proteins are artifically made negative)
What type of gel electrophoresis depends on size and charge Native (protein charge not affected)
Name a factor affecting protein migration apart from size or charge size of pores in buffer
What are the types of microscopy 1. bright field 2. Fluorescence
Which type of microscopy allows tiny structures such as individual proteins within the cell to be viewed Fluorescence
Which type of microscopy is more limited in its resolution Bright field
Which type of microscopy is more simple to use Bright field
Which type of microscopy involves antibodies to tag objects Fluorescence
Name an object that can be viewed under a bright field microscope whole organisms, parts of organism or thin tissues
What does a haemocytometer estimate Total cell count
What technique is able to distinguish between dead cells and viable cells Vital Staining with trypan blue
Explain why only dead cells stain with trypan blue Trypan blue toxic so actively pumped out by living (viable) cells
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
What term is given to cells that can only divide a limited number of times (hayflick) limit primary cell lines
What term is given to cells that can divide infinitely Cancer cells
Why are growth regulators/hormones used in plant tissue To enable explant to divide into callus AND to enable callus to differentiate into plantlet
Why do mammalian cells need foetal bovine serum For cell proliferation
Why do mammalian cells need antibiotics to kill bacteria which grow more readily than mammalian cells
What type of organisms are produced from inoculum microbes
What type of organisms are produced from explants plants
Describe the 5 key stages of mammalian cell sub culture Use proteolytic enzymes to remove mammalian cells from tissue Cells adhere to surface Cells flatten Cells divide to form a monolayer Cells form confluent Process repeats
What is a protoplast plant cell without cell wall
How are different species of plants produced in a lab 2 protoplasts fused together with electric shock & cell wall reformed plant growth regulators then added to divide cells to form callus which differentiate into plantlet
Name a plant hormone/growth regulator auxin or cytokinins
Name a substance a plant needs to grow except plant hormones M & S salts, Macroelements NPK or Microelements Cu, Zn, vitamins, glucose
What two cells fuse to make a hybridomas B lymphocyte with a myeloma cell
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
What substance are the hybridomas placed in and subsequently diluted to ensure one cell per well polyethylene glycol (PEG).
Created by: kyle_academy