Bio II Lab Test 2 Study Guide PDF Fall 2024

General Biology 2 BZE Fall 2024 STUDY GUIDE FOR LAB TEST 2 Date of lab test: Friday, November 29, 10 AM – noon (section 9); 2 -4 pm (section 10) Lab 5 – Fermentation & Respiration  Do you...

General Biology 2 BZE Fall 2024 STUDY GUIDE FOR LAB TEST 2 Date of lab test: Friday, November 29, 10 AM – noon (section 9); 2 -4 pm (section 10) Lab 5 – Fermentation & Respiration  Do you understand the basic differences between the processes of cellular respiration & fermentation, including whether oxygen is required or not, their reactions, and the relative amount of energy that can be made? o Describe the process of cellular respiration, knowing the chemical equation, what conditions it occurs in, location in the cell, and give the ultimate goal. o Briefly, know the steps of cellular respiration and know what is produced at each step. o Describe the process of fermentation, knowing the chemical equation, what conditions it occurs in, location in the cell, and give the ultimate goal. o Do you know the three main pathways that metabolize pyruvate? Which one oxidizes pyruvate? Which ones reduce pyruvate? o Do you know the difference between alcohol and lactic acid fermentation in terms of the steps and products formed?  Can you explain the relevance of the R.Q. ratio (respiratory quotient) in terms of types of substrates being used (carbohydrates or fats) in cellular respiration?  For the pea seedlings demonstration of cellular respiration, do you understand the setup with the soda lime? What would a positive gas pressure indicate? What was the purpose of covering the seedlings with foil?  Are you able to explain all experiments (two) conducted in simple terms using proper vocabulary and ingredients (ie. Pea seedlings, soda lime, cellular respiration, gas pressure sensor (don’t need to explain the software program), yeast, fermentation, glucose, sucrose, maltose, lactose).  Can you explain why the fermentation rates for glucose and sucrose are relatively higher than maltose and lactose? And why did you measure a second maltose fermentation rate after a 15 minute incubation?  The questions in this lab report are at the level of detail you need to know for the test  Do you know these technical/analytical skills: o Given a data set, be able to draw a graph depicting O2 uptake or CO2 emission over time. o Given a graph of fermentation rates or a graph of cellular respiration rates, be able to calculate the rate of O2 uptake or CO2 emission (depending on the scenario). o Calculate the Respiratory Quotient (R.Q.) from a given oxidation reaction, and how it can change given different substrates (sugars, fats, proteins). Lab 6 – Structure & Function of Leaves  Are you able to label and give the function of cells and structures of different leaf cross sections; cuticle, upper epidermis, lower epidermis, palisade mesophyll, spongy mesophyll, vascular tissue, xylem, phloem, stomata, guard cells?  If you know the structural differences between different types of plant leaves, can you explain how they reflect the adaptation of those plants to different environments with respect to: water conservation, gas exchanges & light absorption?  Are you able to identify a mesophyte, xerophyte or hydrophyte leaf in a microscopic cross section by characteristics and identify their components. Page 1 of 7 General Biology 2 BZE Fall 2024  Do you understand the structure and function of stomata: o When are guard cells turgid? Flaccid? How are these states related to opening and closing the pore? o Why do guard cell have chloroplasts? o How do the proton pumps in guard cells affect osmosis and pore-opening and closing?  Do you understand the difference between absorption & reflection of light, & that the light that is absorbed “powers” photosynthesis  Do you understand how paper chromatography separates the pigments (what other molecules can chromatography separate?  What is the stationary phase and mobile phase of chromatography? Why is selecting the appropriate eluent so important to chromatography? What eluent was used in this experiment?  Are you able to explain the experiment in this lab conducted in simple terms using proper vocabulary and ingredients (spinach, chlorophyll, xanthophyll, carotenes, chromatography, absorbance).  Why doesn’t chlorophyll a fluoresce in an intact plant?  Do you know these technical/analytical skills: o Given a completed chromatography paper, be able to calculate Rf values. What does a larger Rf value indicate for a pigment? A smaller Rf value? o Understand how and why the different pigments were separated using the structural difference between chlorophyll a and b as an example. o Can you observe, graph & understand the absorption spectra of the 4 different plant pigments we looked at in the lab. o Are you able to identify a plant pigment based on observing its absorption spectrum? o Given a data set, be able to draw a graph depicting absorbance at different wavelengths. Lab 7 – Bacterial Transformation & Restriction Enzymes Bacterial Transformation: What is a transgenic organism? What was the transgenic organism you created in this lab? created Do you know what a horizontal gene transfer event is, its relationship to bacteria transformation and how it is different from a vertical gene transfer? Be able to define what bacterial plasmids are and what their purpose is in bacteria. Do you understand the function and the relationship between the GFP, beta lactamase (bla) and ara C genes on the pGLO plasmid? What phenotype does the bla gene gave to transformed bacteria? Is this gene constitutively expressed or regulated in the pGLO plasmid? How does the ara C gene regulate the GFP gene? Briefly describe this arabinose operon. o What is meant by a regulatory protein? What role do they play in gene expression? How are they activated or inactivated? Use the Ara C protein as an example. Can you characterize bacterial colony morphology? What is the difference between a bacterial lawn and a bacterial colony? What could alter a bacteria’s morphology? Do you understand the importance of the control treatments that were used in the experiment: Why was the –pGLO in LB plate a positive control for this experiment? Why was the –pGLO in LB + ampicillin plate a negative control experiment? Can you explain from the results of the four bacterial plates why the pGLO plasmid was successfully transformed in this experiment? What does transformation efficiency measure? How would you determine this efficiency measure from the four plates in this experiment? Page 2 of 7 General Biology 2 BZE Fall 2024 What was the purpose of the transformation solution, CaCl2? What was the purpose of the heat shock procedure (ice-heat-ice)? Why was conducting the experiment with aseptic technique (as much as possible) so important? Restriction Enzyme Digest of pGLO: Be able to explain the pGLO restriction digest experiment conducted in simple terms using proper vocabulary and ingredients (ie. DNA extraction, restriction endonucleases, restriction sites, electrophoresis, agar, EcoRV, HindIII, loading dye). Can you explain the use of restriction endonucleases by bacteria to destroy viral DNA and how they protect their own DNA by methylation? Can you describe what a recognition site is and restriction fragments? Can you explain what are “sticky ends” and “blunt ends” when a restriction site is cut by its enzyme? Are you able to describe how restriction endonucleases work and be able to predict how often DNA will be cut by different restriction endonucleases based on the restriction site? When preparing the pGLO plasmid for digestion with various restriction enzymes, why was a buffer added? Why were the restriction enzyme(s) + pGLO plasmid samples placed in a 37oC bath and then heat- shocked at 65oC? Describe gel electrophoresis and what it was used for in this experiment: o What did this procedure separate? o What is the unit of size for DNA? o What were the two purposes of adding the 6X loading dye to each digested sample before loading them onto a gel? (How do you know when to stop o What is the purpose of loading a lane in the gel with DNA ladder? o What was the purpose of loading a lane in the gel with uncut plasmid? Why were there two bands in this lane? o What is the overall charge of DNA (positive or negative)? o Why would DNA bands move towards the positively charged electrode in a gel when there is a current running through the gel? o Why are smaller DNA bands migrating further down the gel than larger DNA bands? o What happens if you don’t pay attention to the gel and leave it running for hours? Where do the DNA bands go? Do they stay in the gel? If you are given a completed gel, are you able to read the separation of DNA fragments, knowing which fragments are smaller and which are larger? If you are given a completed gel, are you able to determine how many restriction sites are present in a plasmid for a specific restriction enzyme? Are you able to determine which restriction sites belong to which restriction enzyme if you are given a completed gel and a plasmid map? Page 3 of 7