Review for Lab Competency Exam

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Review for Lab Competency Exam

• Recitation

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Admin

• Sign up for specific time / date
• If late, your TA has the discretion to let you
  come in (miss stations) or not (automatic 0)
• Need accommodation must reschedule 1 week in
  advance
• Make ups reschedule by contacting Tom (432-1316
  ext 137) immediately

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Hints for Studying

• Practice timing yourself doing the different
  stations (read through study guide and imagine
  yourself doing the work)
• Practice graphing all the data we have collected
  during the semester, so you can quickly remember
  which variable goes on each axis and pick the
  right paper remember you must print the graph
  before time runs out make sure you staple this
  graph to your test since if you lose it you will
  not be allowed to make another one every time
  you do a graph, verify you can make it and print
  it in less than 8 minutes
• Make sure you can identify TC vs TD devices and
  you know how to convert mL to Fl or Fl to mL
  know which tools are the most accurate and how to
  use the P20 and P1000 properly
• Review and be able to look at cells to determine
  whether solutions are hypertonic
  /hypotonic/isotonic and use the correct terms to
  describe what is happening to the cell and which
  way water moves

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Logistics
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Results Bacterial Transformation
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Results Bacterial Transformation
Conclusion - Green unknown is pAMP and brown
unknown is pKAN
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Station 4 Gel electrophoresis, page 20

• Separating out DNA fragments in 0.8 agarose due
  to differences in size (base pairs) and shape

• DNA must have a negative
• charge
• Be soluble in solution
• Different shapes (linear,
• supercoiled, nicked relaxed)

Cathode

• larger linear fragments move
• less distance than smaller linear
• fragments

Anode
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Maps
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Restriction Digest Gel electrophoresis
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Result Restriction Digest Gel electrophoresis
Lambda DNA Hind III Linear DNA
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Result Restriction Digest Gel electrophoresis
Lane 1 2 3 4 5
6 7 8 9 10
Which gel pKAN?
Which lane(s) marker?
A
D
F
Which gel pAMP?
Which band supercoiled? nicked, linear?
E
B
G
C
Which U? D? S?
Gel 2
Gel 1
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Safety Disposal

• NFP symbol (red fire yellow reactivity blue
  health white special
• Scale 0 least hazard to 4 highest hazard

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Safety Disposal

• Solid biohazardous waste can
• Broken glass can
• Solid chemical waste box
• 5-gallon carboy (liquid chemical wastes)
• Trash can
• 1-liter beaker
• Sharps biological container

• Office of Radiation, Chemical Biological Safety
  (ORCBS)
• Material Safety Data Sheet (MSDS)

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Techniques Tools

• Streaking
• 1. Isolation
• 2. Confluency
• Spectrophotometry
• Centrifgation
• Gel electrophoresis
• SDS-PAGE (protein)
• Agarose (DNA)
• Microscopy
• Measuring tools
• Paper chromatography
• Respirometers
• Colloid
• Biochemical tests

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Graphing

• Standard Curves
• Optimal Curves
• Time Course Curves
• Bar graph

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Spec Graph
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BS111L Recitation

• Graphing (page 33 34)
• Standard curve

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Protein Concentration Biological Molecules lab

• 4. Results

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Colloid Analysis

• 4. Results

y-axis can label as Molecular Weight (g/mole)
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BS111L Recitation

• Graphing (page 33 34)
• 2. Optimal curve

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Enzymes Environmental Analysis

• 4. Results

Acid hydrolysis
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Enzymes Environmental Analysis

• 4. Results

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BS111L Recitation

• Graphing (page 33 34)
• 3. Time course curve

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Enzymes Environmental Analysis

• 4. Results

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Glycolysis coupled to Respiration

• 4. Results

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Glycolysis coupled to Respiration

• 4. Results

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Glycolysis coupled to Fermentation

• 4. Results

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Station 5 Centrifugation, page 21
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Station 5 Centrifugation, page 21
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Station 3 Measuring page 17
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Station 3 Measuring page 17
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Station 3 Measuring page 17
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Station 2 Steaking page 40

• 4 tools for confluency
• 95 ethanol in a beaker
• Incubator
• Bunsen burner
• Bacteria
• Hockey stick
• Agar in petri dish
• Pasteur pipette bulb
• 1 L waste beaker

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Station 2 Steaking page 40
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Dilution

• Manipulate the formulas (How set up dilution if
  you want to make 10 mls for each dilution? )
• C1V1 C2V2 and V2 V1 Vd
• C1_V1_ C2
• V2
• __V1__ __1___ __V1__
• V2 2 V1 Vd

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Microscopy, page 32
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Microscopy, page 32
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Station 4 Gel electrophoresis, page 20

• Separating out different sizes (mass or molecular
  weight) of proteins in SDS-PAGE (polyacrylamide
  gel)

• proteins must have a negative charge
• Be soluble in solution
• Be denatured (linearizes)

Cathode
- Smaller protein chains move farther distances
than larger protein chains
Anode
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SDS-PAGE
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Protein Fingerprinting Analysis

• Results
• Between most similar or least similar related
  species calculate percent commonality
• Calculate in Common between two species

number of proteins in common between both
species ------------------------------------------
---------------------------------
total number of different proteins present in
both species
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Protein Fingerprinting Analysis

• Results Calculate in Common between most
  similar related species salmon cod

AM Fl Co Sa K Ca Sc
MHC A T MLC MLC
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Protein Fingerprinting Analysis

• Results Calculate in Common between to each
  group of species

Notice scallop similar to other marine fish
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Protein Fingerprinting Analysis

• 4. Results Determining sizes of unknown proteins?

Catfish Protein 26 mm So how Big?
catfish
50 kD
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Glycolysis coupled to fermentation
LAB

• measure air/gas level of each small test tube
• _at_ 0min 60min
• 37C bath
• check after 30min
• fill out table 1
• calculate CO2 production
• in mm
• volume of CO2 production
• use 10mm 1ml CO2
• Create BAR / COLUMN GRAPH

label
1
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Lab Cellular / Organismal Respiration

• cotton protection and diffusible
• KOH corrosive / absorb CO2
• measuring pipette O2 consumption
• clamp to close tube - after equilibration!!!
• dye for measuring pipette
• visualize O2 consumption
• loading dye
• reading every minute over 20 minutes
• weigh the tube before after loading with
  organism

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Lab Cellular / Organismal Respiration
CO2 KOH
PEA SEED
O2
Decrease of pressure in the tube compared to the
normal air atmosphere outside the tube!!!!!
Movement of the dye
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Photosynthesis
Respirometer used to measure oxygen production
from the plant doing the light reactions
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Biological Molecules (Parts II IV)
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(No Transcript)
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Part II Differentially permeable membrane
What happens to weight of bag and why?
?
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Cells in a
Hypertonic solution high conc solute / low
solvent (H2O) conc.
Hypotonic solution low conc solute / high
solvent (H2O) conc.
Isotonic solution same conc solute / same
solvent (H2O) conc.
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Osmosis
Which beaker(s) will water move into the
bag? Which beaker(s) is(are) isotonic to the
bag? Which bag(s) is(are) hypertonic to the bag?
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Which direction does the water move?
Is the bag hypertonic or hypotonic or isotonic
compared to the beaker water?
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Hints When Taking the LCE

• If have extra time at a station, then start
  reading ahead for what you are supposed to do at
  the next station (planning)
• When get to a station, read through all the
  questions and work on the questions which require
  letter answers from the station first then go
  back and do technique work so if time runs out
• Decide which station makes you the most nervous
  and avoid starting the LCE at that location
  either start right after it so it is your last
  station or decide which station you would like to
  start at and get there first
• Decide ahead of time if you are willing to give
  up doing certain techniques and losing some
  points so you can stay calm during the exam
• Not looking around and watching other people you
  focus on what you are doing and your exam