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Fall 2009
Instructor: Dr. Mary Ogilvie
Office: S203F
Phone #: 321-3437
Email: mogilvie@cbu.edu
Grade based on: 2 exams ------------- 100 pts. apiece
~12 Homework assign. --- 10 pts. apiece
Total ---------------320 pts.
Homework Assignments: Can be done in groups of two unless otherwise stated. This means both people must work on the homework. Assignments are due the Tues. following lab.
Make-up Exams: Should you not be able to attend lab the day of an exam, please call me as soon as possible. You will be asked to present a Drs. note.
Grading Scale: A = 90-100 D = 60-69
B = 80-89 F = <60
C = 70-79
Course Goals:
1. To develop hands-on experience with such techniques as column chromatography, fluorescence microscopy, electrophoresis, spectrophotometric analysis and PCR.
2. To understand the importance of positive and negative controls in experimental design and implementation.
3. To be able to critically evaluate and interpret experimental results.
4. To gain greater familiarity with red blood cells (RBCs) and platelets by studying their structure and function.
5. To understand how cells such as RBCs, platelets, transformed mammalian cells, and bacterial cells can be used as tools to study cellular processes, some unrelated to the cell itself.
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Date Experiments Exercise
Jan. 15 Introduction: Pouring Sepharose 4B columns
Jan. 22 Purification of Venom Lectins by Affinity Chromatography
-Pervaporate by next lab.
Lectin and Venom Hemagglutination.....Hmwk
Jan.
29 Protein Determination of Lectin with BioRad Reagent
Run Excel to obtain standard curve ----- Hmwk
Do samples for SDS gels (2 red. and 1 NR gel; each group makes 3 samples)
Lectin Hemagglutination Assays-----Hmwk
Run Lectins onSDS gels
Feb.12 Start SDS PAGE for Western Blot with Lectins (Use samples I provide) Molecular Wt. Determination of Lectins on SDS PAGE------- HMWK
Start Immunofluorescence of RBCs + Lectins-----
Hmwk
Feb. 19 Finish and Evaluate Western blots
-Summarize & explain relationship to SDS gels (last week)-- Hmwk.
Coat Adhesion Plates with Plasma
Finish Immunofluorescence
Feb. 26 Cellulose Acetate Electophoresis of Lectins
Begin Platelet Adhesion Assays
Run Adhesion data on Excel and answer homework questions -- Hmwk
March 11 Spring Break
March 18 EXAM I (Covers all labs done thus far, except platelet adhesion assays)
DNA Forensics Digestion with Hind III and Digestion of Lambda DNA
Review Mid-term Exams
Run and Evaluate Forensics and Lambda DNA gels----- Hmwk.
April 1 Purification of GFP by Column
Chromatography
Buccal Cell DNA Extraction, Polymerase Chain Reaction with Alu
Evaluate and summarize Transformation frequency -- Hmwk
April 8 EASTER BREAK
April 15 pGLO Purification
with Qiagen Maxi Kit
Agarose gel with Amplified Alu
-Evaluate & summarize Alu gel ----- Hmwk
April 22 Spectrophotometric Analysis of pGLO and GFP
-Evaluation and Summary of Spec. analysis ---Hmwk
Digestion of pGLO with restriction endonucleases
April 29 Agarose Gel Electrophoresis of pGLO
-Evaluate and Summarize pGLO Gel-- Hmwk
Lab Review
Final Exam either Friday April 30 or as scheduled during Exam week.
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