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Advanced Higher Bio
Unit 1 lab techniques
| Question | Answer |
|---|---|
| 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
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