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Syllabus | Lab Schedule | Lab Report Protocol | Lab Outline 1 | Final Lab Outline
Biology 111L LAB
Fall 2004
Instructor: Dr. Mary Ogilvie
Phone: 321 - 3437
email: mogilvie@cbu.edu
Office Hrs.: MWF 1:00 4:00
Thurs. 9:30 10:30
Corequisite: Biol. 111
Books Required: 1.Biology Laboratory Manual 6th
ed., Warren D. Dolphin
2. Photo Atlas for Biology, 2nd ed., Perry and Morton
Grading: 1 mid semester exam/practical (100 pts)
1 comprehensive final exam (150 pts)
2 lab reports (50 pts. each)
~10 weekly quizzes (20 pts. each)
Total: ~550 pts.
If you attend all of the labs, you may drop your two lowest
quiz grades. If you miss one lab, you may drop your single, lowest
quiz grade.
Attendance: For any missed labs, a Drs. note must be
presented as soon as possible. If you are ill, please call and
let me know prior to lab. For each unexcused lab missed, 10 pts.
will be deducted from your overall lab grade.
Quizzes will be given at the start of the period. If you are late
and miss a quiz, there will be no make-up quiz. Makeup quizzes
will not be given regardless of whether an absence was excused
or not.
Goals:
- To become proficient using such tools as microscopes, pipets,
spectrophotometers, micropipetters, and manometers.
- To be able to distinguish and identify cells from organisms
within the five biological kingdoms.
- To develop an appreciation of scientific enquiry and of the
importance of developing and using controls in an experimental
system.
- To learn the anatomic and functional differences characteristic
of organisms within the Kingdoms Fungi, Protista and Prokaryotae.
- To be able to use basic statistical tools for the interpretation
of experimental data.
LAB REPORT PROTOCOL
PLAGIARISM: Occasionally students will use passages, verbatim, (often without evil intent) from other sources (books, jourals, etc.) in a report. This is PLAGIARISM and is considered a form of cheating. The only acceptable way to use another source is to put the passage in your own words and the include it in the citation section (see below). You will receive an F on your report if you do this.
You will be asked to write two lab reports. Choose between the first three labs listed with an asterisk. This report will be due 2 weeks after the lab exercise. The 2nd report must be a fruit fly report and will be due near the end of the semester (see last lab with an asterisk).
I. Abstract ---- This is a summary of the entire experiment. The Abstract includes a brief summary of your results. It is condensed into one paragraph.
II. Introduction ----- Includes any important facts pertinent to understanding the experimental parameters. Much of the information given in pre-lab would be found here.
III. Hypothesis------ A testable statement that is one sentence stating predicting the results. Your results may not always support the prediction. If it does not then include in the Discussion (below) why your results did not support your hypothesis.
III. Methods ------ This section is a brief summary of how the experiment was run. No quantities of materials are necessary here.
IV. Results ------ A verbal explanation of your results is provided here. Include any data in tables or graphs that are clearly labeled. Any calculations, which apply, are included here also. GRAPHS MUST BE DONE IN EXCEL. Always explain tables, graphs, etc. No commentary about your results is needed here. Just the Facts!
V. Discussion ------ Give your interpretation of the
results. Why do think your experimental results were good or bad?
How did your results align with the information given in your
Introduction? If your results were poor, give some idea as to
why they were less than perfect. Be honest. You will not be penalized
for poor results. I am most interested in knowing that you understand
the experiment. This section requires several "meaty"
paragraphs.
-Avoid editorializing, such as making statements that begin with,
"I feel..." or "All in all the experiment went
well." or "The experiment went as expected."
VI. Citations------ Any sources included (in your own words) in your reports should be placed at the end of the report. Citations will be listed numerically in order of their appearance in the paper. In the text, references should be cited by putting a number (in superscript) at the end of the paragraph containing the citations. I will hand out a sheet showing you how this is done.
Extra tips:
1. Paper must be written in 3rd person, passive voice, past tense.
2. . Each time a new topic is addressed, begin a new paragraph,
each of which should have a topic sentence. (If you don't know
what this is, ask me.)
3. Use BOLD headings and Roman numerals to delineate each
section of the report.
4. Reports must be typewritten and at least 3 pages.
5. Please write reports in groups of 2 whenever possible.
Lab Schedule
Week of Exercises Topics for Investigation
8/31 2, 3 Microscopy; Cell Structure
9/7 4 Quantitative Lab Tech.; Statistics
*9/14 6 CellularTransport Mechanisms
*9/21 5 Properties of Enzymes
*9/28 7, 25 Cell Respiration, Chlorophyll Absorption Spectrum
10/5 8 Cytogenetics: Chromosomes and Mitosis
10/ 12 Exam I
10/19 FALL BREAK
10/26 10 Start Fruit Fly Crosses 9 Cytogenetics Meiosis & Crossing-Over
*11/2 10 Mendelian Genetics I: Fruit flies,
Human inheritance
11/9 11 Experiments with DNA
11/16 12 Microevolutionary events
11/23 13,14 Bacteria and Protista
11/30 17 Fungi
12/7 FINAL LAB EXAM Biology 111 Lab Outline I Exercise 2 Microscopy A. Magnification (Know metric units) B. Resolution 1. Naked eye 2. Compound 'scope 3. Electron 'scope C. Contrast D. Handling microscope E. Know parts of microscope: 1. Ocular (10x) 2. Objective lenses -Know names of each *Total magnification = Ocular x objective 3. Stage 4. Base and arm 5. Coarse and Fine adjust. knobs 6. Mechanical stage 7. Turret 8. Substage condenser 9. Diaphragm F. Field of View -estimate with graph paper G. Compass slides -Image is upside down and reversed H. Different types of Microscopes 1. Compound Microscope 2. Dissecting Microscope 2. Electron Microscopes a) Transmission E.M. b) Scanning E.M. Exercise 3 Cell Structure Reflects Function A. Prokaryotes 1. Characteristics? 2. Lactobacillus in yogurt a) Digests lactose b) Lactic acid drops pH c) Curdles milk 3. Anabaena a) Blue-green algae b) Picture p. 194 B. Wet Mount C. Eukaryotes 1. Characteristics? 2. Kingdom Protista -Paramecium 3. Kingdom Fungi -Yeast budding 4. Kingdom Plantae -Epidermal cells of onion 5. Kingdom Animalia a) Spinal Cord Neurons b) Buccal smear Exercise 4 Quantitative Lab Techniques I. Pipetting A. Types of Pipettes 1. Serological 2. Mohr 3. Volumetric B.Technique 1. Meniscus 2. No fingers on tip C. Pipetting Experiment 1. Taring 2. % Error (Pos. and Neg.) 3. Calculate density D. Simple Statistics 1. Mean 2. Range 3. Variance 4. Standard Deviation II. Spectroscopy A. Spectrophotometer 1.How does it work? 2. Two measurements om spec. a) Absorbance b) % Transmittance B. Use of Spec. 1. What is a blank? 2. How do you zero machine? 3. How do you set wavelength? C. Optimal Wavelength 1. Test Meth. Blue at different l 2. Plot Abs. vs. Wavelength a) Independent var. = Y-axis b) Dependent var. = X-axis D. Standard Curve 1. Used to determine unknown concentration 2. Use optimal l 3. Plot concentration vs. absorbance 4. Line of best fit 5. Find absorbance of unknown on line Exercise 5 Properties of Enzymes A. Enzymes 1. Modify substrate -bind via active site 2. Act as catalysts 3. Peroxidase Test a) 2H2O2---¦ 2H2O + O2 b) Guaiacol turns brown in presence of O2 -Amt. of brown proportional to enzyme activity B. Inhibition of Peroxidase 1. Hydroxylamine a) Binds active site of enzyme b) Prevents binding of substrate 2. Boiling = Denatures enzyme 3. pH extremes = interfere with charge interaction between E and S 4. Temperature extremes C. What Blank Was Used? Exercise 6 Cellular Transport Mechanisms A. Brownian Movement 1. Carmine dye in detergent 2. Vibrating motion due to molecular collisions B. Diffusion 1. Agar Diffusion a) Porous material b) Apply ferrous sulfate and potassium ferricyanate to 2 wells c) Lgr. molecules move slower than smaller d) Formation of Prussian Blue in middle 2. Dialysis a) Large molecules cannot pass out of tubing pores b) Tests: i. Cl- = Use AgNO3 -Get white ppt.= AgCl ii. SO42- = Use BaCl -Get white ppt.= BaSO4 iii. Albumin = Use BioRad -Get blue color change iv. starch = Use I2KI -Dark Brown or purple c) Identify Pos. & Neg. controls 3. HCl + NH4OH in glass tube a) Precipitate formation = NH4Cl b) Precipitate forms closer to side with lgr molecules -Why? C. Osmosis 1. Paremecium (protozoan) a) Lives in hypotonic environ. b) Collects water & expels H2O c) H2O expulsion vacuoles d) No lysis 2. Osmometer a) Water moved into thistle tube b) Crossed semi-permeable membrane c) Why did it move in this direction? 3. Dialysis -What about this experiment demonstrated osmosis? Exercise 7 Cellular Respiration I. Glycolysis and Fermentation A. Be familiar with general scheme of both B. Ethanol Fermentation 1. Karo + H2O + Yeast a) Karo is carbon source b) Yeast is source of enzymes 2. CO2 release a) Bubbles released b) Formation of BaCO3 -How? 3. Distillation to isolate ethanol -Principle behing this? 4. Iodoform test a) Test for ethanol b) What were the positive and negative controls? c) How performed? C. Aerobic Respiration 1. Peas incubated in sucrose a) Plus azide -Inhibitor of electron transport b) Minus azide 2. Peas placed in manometer -What is manometer? 3. Purpose of KOH? 4. Measure O2 consumption a) Watch dye move in pipet over time b) Why does O2 move inward? 5. Purpose of thermobar? 6. How are data points calculated? 7. Graph a) Two lines of best fit (plus and minus azide) b) Why is slope necessary? c) Why is weight of peas necessary? Exercise 8 Mitosis A. Animal and Plant Mitosis 1. Know differences between: a) Plant = Onion root tip i. Cell wall ii. Phragmoplast at Telophase b) Whitefish blastula (embryo) i. No cell wall ii. Cleavage furrough 2. Know phases a) Prophase i. Chromosome condensation ii. Disappearance of nuclear membrane and nucleolus iii. Formation of spindle b) Metaphase i. Chrom. most condensed ii. Chrom.in middle of spindle c) Anaphase i. Chromatids separate ii. Chromatids move to poles d) Telophase i. Chromosomes more diffuse ii. Reappearance of nuclear membrane and nucleolus iii. Formation of phragmoplast or cleavage furrough B. Preparation of Onion Root tip slides 1. Fixative a) Denatures proteins b) Extract lipids from membranes 2. HCl a) Hydrolysis of cell wall proteins b) Some hydrolysis of chromosomal proteins 3. Feulgen stain = stains chromatin Exercise 9 Meiosis and Crossing over I. Spermatogenesis A. General diagram B. Slide of mammalian testis 1. Seminiferous tubules 2. Developing sperm II. Oogenesis A. General diagram B. Slide of mammalian ovary 1. Graafian follicles 2. Follicle cells 3. ovulation C. Ascaris oocytes and zygotes 1. Parasitic roundworm -4 chromosomes 2. Meiosis in egg occurs after fertilization 3. Identify: a) Sperm b) 1o oocyte c) 2o oocyte d) polar body e) Pronuclei f) Shell III. Crossing over in Sordaria A. Know life cycle B. How was cross between two strains performed? C. Be able to diagram: 1. No cross over asci 2. Crossing over between chromatids of two different strains D. Identify: 1. Perithecium 2. Ascus 3. Ascospores 4. Hyphae
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Study Outline for Final Exam
UPDATED 11/28/01
Remember, the exam will be comprehensive!!!!!!
Exercise 9 Meiosis and Crossing over
I. Spermatogenesis
A. General diagram
B. Slide of mammalian testis
1. Seminiferous tubules
2. Developing sperm
II. Oogenesis
A. General diagram
B. Slide of mammalian ovary
1. Graafian follicles
2. Follicle cells
3. ovulation
C. Ascaris oocytes and zygotes
1. Parasitic roundworm
-4 chromosomes
2. Meiosis in egg occurs after fertilization
3. Identify:
a) Sperm b) 1o oocyte
c) 2o oocyte d) polar body
e) Pronuclei f) Shell
III. Crossing over in Sordaria
A. Know life cycle
B. How was cross between two strains performed?
C. Be able to diagram:
1. No crossing over asci
2. Crossing over between chromatids of two different strains D. Identify:
1. Perithecium
2. Ascus
3. Ascospores
4. Hyphae
Exercise 10 Genetics
I. Human Genetics
A. List of Human Genetic Traits
-See handout
B. Blood Typing
1. ABO Blood groups
a) A, B, AB have antigens
b) O has no ABO antigens
c) Type A individuals make Abs to Type B and AB
d) Type B individuals make Abs to Type A and AB
e) Type O
i. Universal donor
ii. Make Abs to A, B and AB
f) Type AB
i.Universal recipient
ii. Make no Abs
2. Rh blood groups
a) Rh + have antigen
b) Rh+ = 95% of population
3. Blood serum
a) Contains Abs against A and/or B Ags
b) Clumping due to cross-linking by Abs
c) How were our tests interpreted?
II. Drosophila Genetics
A. Know life cycle
1. Egg 2. Larva
3. Pupa 4. Adult
B. Know reasons for using fruitflies for experiments
C. Distinguish the Two Sexes
D. Crossing fruit flies
1. White eye
a) sex-linked mutation of eye color
b) Recessive to red
c) males hemizygous
2. Be able to diagram these crosses
from parents to F2's
a) White male x Red female
b) Red male x White female
3. Know expected F2 ratio
E. Chi Square = Statistical Tool
1. Null Hypothesis, define.
2. Be able to do a Chi Square analysis and determine:
a) Confidence level
b) Degrees of freedom
c) Whether to accept or reject Null
III. Corn
-Be able to do monohybrid and dihybrid crosses with corn
Exercise 11 Molecular Genetics
I. DNA Extraction
A. Isolation from onion
B. Know why following used:
1. Woolite
2. NaCl
3. Incubation at 60oC
4. Ethanol
C. Why should you do the following?
1. Keep solution cool
2. Pipet ethanol slowly
II. DNA Agarose Electrophoresis
A. Agarose
1. Porous matrix
2. Microwave to dissolve
3. Cool to harden
B. Separation by size
1. Large molecules move slower
2. Small molecules found farthest from wells
C. Technical Details
1. Add sample buffer to DNA.Why?
2. Add DNA to wells
3. Turn on electric current
4. Stain and destainWhy?
Exercise 13 Microevolution
I. Hardy Weinberg Principle
A. Hardy-Weinberg Equation
a) p2 + 2pq + q2 = 1
b) p+q = 1
c) Means of determining allele frequencies in a population
Didn't do these fall of 2001:
B. Bean Exercise
1. Accounting Method to determine frequencies
2. Beans represented mating pairs
3. Why was each mating event multiplied by 4?
4. Gene freq. should not change much from generation to generation
C. You Be the Predator
1. Dark backgroud
a) Light beans picked off
b) Freq. of light allele decreases
c) Freq. of dark allele increases
2. Light backgroud
a) Dark beans picked off
b) Freq. of dark allele decreases
c) Freq. of light allele increases
3. Be able to interpret graph
II. Serrartia experiment
1. Serratia (bacterium)
a) Orange at room temp.
b) White at 37oC
-Can't make pigment at this temp.
2. Induce mutation with UV (mutagen)
3. White colonies at room temp. indicate mutation
4. Calculate mutation rate
5. Why did we use hockey sticks?
Didn't do these fall of 2001:
Exercise 14 Bacteria
I. Bacterial Characteristics
1. Prokaryotic
2. Mostly heterotrophs
3. Identify by shape:
a) Cocci
b) Bacilli
c) Spirilla
4. Identify by Gram stain
a) Identify reagents b) Gram (+)
i. Peptidoglycan layer stained purple
ii. cell wall not dissolved by ethanol
c) Gram (-)
i. Red
ii. Lipopolysaccharide dissolved by ethanol
iii. Peptidoglycan stained by counterstain (safranin)
II. Bacterial Organisms
1. Serratia
2. Bacillus
3. Cyanobacteria
a) Blue-green algae
b) Contain chlorophyll
-autotrophs
c) Gleocapsa = 4 or more cells surrounded by sheath
Exercise 14 Kingdom Protista
I. Characteristics
1. Mstly unicellular
2. some colonial
3. Eukaryotes
Didn't cover in fall 2001:
II. Photosynthetic Protists
A.Phylum Dinoflagellata
Ceratium
1. Spikes
2. Two flagella
B. Phylum Euglenophyta
Euglena
1. Flagellum
2. Heterotrophic and photosynthetic
Responsible for in 2001:
C. Spirogyra
a) Spiral chloroplasts
b) Conjugation = sexual reprod.
D. Diatoms
a) Covered by two valves
-Silica
b) Diatomaceous earth = layers of dead diatoms
c) Used for swimming pool filters and toothpaste
III. Heterotrophic Protists
A. Amoeba
1. Movement by pseudopodia
2. Phagocytosis
3. Entamoeba histolytica
a) Unwashed fruits and vegetables
b) Causes Amoebic dysentery
B.. Phylum Ciliophora
Paramecium
1. Cilia
2. Macronucleus
3. Oral groove
4. Contractile vacuoles
D. Trypanosoma
a) Blood parasite
b) Tse tse fly = vector
c) African sleeping sickness
Exercise 17 Kingdom Fungi
I. Characteristics
1. Hyphae, Mycelium
2. Heterotophs and saprobes
a) Enzymes break down nutrients followed by absorption
b) Decomposers
3. Reproduction
a) Sexual by fusion of 1N hyphae
b) Asexual by germination of 1N spores
II. Phylum Zygomycota
Rhizopus
1. Bread mold
2. Hyphal fusion = sexual reproduction
-Zygospores produced
3. Sporangia contain asexual spores
III. Phylum Ascomycota
1. Peziza
a) Cup fungi
b) Asci in ascocarp
-ascospores
2. Sordaria
3. Edible morel
4. Why are all of these called sac fungi?
IV. Phylum Basidiomycota
1. Coprinus
a) Mushroom b) Cap, stem, gills
c) Basidiospores in Basidia
d) Basidiocarp e) Why called club fungi?
2. Bracket fungi
3. Puffballs
V. Lichens
1. Umbilicaria
a) Sac fungus + green algae
b) Algae on outside
c) Extremely hearty. Why?
2. Dried specimens on branch
a) Crustose
b) Foliose
c) Fruiticose
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