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31 Cards in this Set
- Front
- Back
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Light Microscope Parts |
1. Light source 2. Substage condenser 3. Objective lens 4. Ocular lens 5. Eye lens system 6. Focusing knob |
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Iris Diaphragm use |
Controls amount of light passes through Substage condenser |
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Phase-contrast (P-C) microscopy |
converts different refractive index into different intensity; light is hitting specimen at different angles to cause variations in brightness and gives depth to the image |
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Transmission electron microscopes (TEMs) |
form images using electrons that are transmitted (pass) through a specimen |
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Scanning electron microscopes (SEMs) |
utilize electrons that have bounced off the surface of the specimen |
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Reasons to use TEMs and SEMs |
1. Disables proteolytic enzymes 2. Prevent extrinsic damage 3. Increase strength/ stability of sample |
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Differential centrifugation |
a technique used to isolate a particular organelle in bulk so that its function can be studied or so that an enzyme can be isolated from it |
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Ways to Disrupt a Cell |
1. Cryopulverization 2. Detergent Lysis 3. Pressure 4. 'Cell Bomb' 5. Sonication 6. Bead Metal 7. Enzymatic |
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Protein Purification Steps and Purification uses |
Detection--> Quantification --> Purification Purification: 1. Research 2. Industry 3. Theraputic |
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Liquid Chromatography |
the term for a variety of techniques in which a mixture of dissolved components is fractionated as it moves through a porous matrix 2 Phases: 1. Mobile Phase 2. Immobile Phase |
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Ion-Exchange Chromatography (IEC) |
Opposite charge of protein Add sample, elute based on [pH/ salt] to release ion, lower pH to protonate anion, increase salt to outcompete |
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Gel Filtration Chromatography |
Separates proteins (or nucleic acids) primarily on the basis of their effective size Beads with pores of defined size Smaller enter and have longer path because of porous gel Larger bypass and get through faster |
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Affinity Chromatography |
Takes advantage of unique structural properties of desired protein; can specifically withdraw and interact with the desired protein and leave all others behind Near total purification of desired protein in single step |
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Electrophoresis |
techniques depending on the ability of charge molecules to migrate if placed in electric field; widely used technique to fractionate proteins >uses PAGE (polyacrylamide gel electrophoresis) electric current applied to separate molecules, based on molecular weight |
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SDS-PAGE |
done in the presence of (-) charged detergent Anionic detergent coats proteins - linearizes them and imparts (-) charge |
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2D-PAGE |
separates based on 2 different properties: 1. Isoelectric point 2. Then by size |
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Proteomics |
Analyze expression and identify individual proteins |
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Polymerase Chain Reaction (PCR) |
used to amplify, cheaply and readily specify DNA regions without the need for bacterial cells uses reverse transcriptase, and RNA templates |
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Spectrophotometry |
measures amount of light of a specific wavelength absorbed by a solution of protein or nucleic acid More protein = more light absorbed = less hitting photocell (detector) |
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How many milli Moles in one Mole? |
1000 mM = 1 M |
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How many milli liters in a Liter? |
1000 mL = 1 L |
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How many micro liters in a milli liter? |
1000 uL = 1 mL |
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What order do you add the dizaocoupling reagents in the two experiments? |
1 mL 1% sulfanilic acid -- 20% HCl 1 mL 5 mM NED(N-1-naphthylenthylenediamine) |
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What is NED used? |
the Nitrate yields a color development; the colors intensity depedns on the amount of NaNO_2 |
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What was the spectrometer detecting? |
the absorbance of nitrite in the solution |
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Pseudomonas use a series of proteins to convert _____ to _______ |
Nitrate (NO_3) --> Nitrite (NO_2) + H_2O |
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What are the three types of Pseudomonas we used in the lab? |
PAO1 -- original strain SS24 -- has a NAP gene inhibited SS304 -- has a NAP gene inhibited |
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What process was used in the experiment of Pseudomonas? |
Periplasmic nitrate reductase |
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What toxic compound was used in the Pseudomonas experiment, that when vortexed oxidized and stopped the reaction? |
Methyl viologen (was added LAST) |
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What is the difference between Dissimilatory and Assimilatory? |
Dissimilatory [NO_2 --> N_2O --> N_2] >>No electron required; Absence of O_2 Assimilatory [NO_3 --> NO_2 --> NH_4] >What we tested in the lab |
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What is a supernatant? |
The supernatant was the liquid and other materials left in the cuvette after centrifuging; everything BUT the cells in the bottom!!! |
