Department of Biology

Biology 4417/5417 - Molecular Biology

Course Description


Prerequisites: 19 hours in biology including both Biol. 2401 (Microbiology) and Biol. 3300 (Genetics); Chemistry 1401 or 1403. Successfully completed either Biol. 3400 or Biol. 4401 is strongly encouraged. Graduate standing is required if enrolled in #5417. A study of molecular biology theory and practice. Emphasis is on the study of model systems to understand the current approaches and laboratory techniques necessary to answer basic questions in current molecular biology. If taken for graduate credit, the prerequisites also include a BS in Biology or permission of the instructor. Graduate students will be expected also to complete a mini-proposal (NIH/NSF style) which will be defended orally.

TEXTBOOK: Molecular Cell Biology. Lodish et al 4th edition Wh Freeman and Co. 1999

Lecture Syllabus
Introduction
1) DNA Replication/ Organization
2) RNA Transcription/Translation Molecular Biological Techniques
3) Vectors
4) Enzymes used in Molecular Cloning
........................1. Gel Electrophoresis
........................2. cDNA/ Genomic Libraries
........................3. Library Screening
........................4. DNA sequencing
........................5. PCR
........................6. Mutagenesis
Exam #1
Selected Topics in Molecular Biology

...1.Expression of cloned Genes I
...2.Expression of cloned Genes II
...3.Detection of Gene Products I
...4.Detection of Gene Products II
...5.Transcription Regulation I
...6.Transcription Regulation II
...7.Transcription Regulation III
...8.Transcription Regulation IV
...9.Transcription Regulation V
..10. Post-Transcriptional/Translational Regulation
Exam #2
..11. Cell Cycle
..12. Cell Cycle
..13. Programmed Cell Death
..14. Oncogenes I
..15. Oncogenes II
..16. Oncogenes III
..17. Gene Therapy I (Genes in Medicine)
..18. Gene Therapy II (Genes in Medicine)
FINAL Exam

Laboratory Syllabus
Week 1/2
Lab Safety / The Big Picture Tour of the BUSH empire
.........Media Prep/Sterilization/Bacterial Stocks/Tips
.........Autoclave Prep Solutions (TAE / TBE etc.)
.........Pour Gels (Agarose)
........ Pipette Practice
........ Agarose gel electrophoresis. Molecular Weight Estimation
.........Bacterial transformation/plating on LB-Amp plates

Week 3-5
DNA miniprep procedures
.........Isolate DNA and Agarose gel electrophoresis
.........Restriction Digestions
.........Principles of PCR
.........Restriction digest of your miniprepDNA
.........Electrophoresis

START REAL EXPERIMENTS FOR THE BIG PROJECT

Week 5/6
Setup RAB PCR - TM discussion
.........Variables of PCR
.........Run Gel of PCR Rxn
.........Setup Ligation
.........Principles of Vector-Insert Ligations

Week 6/7
Transformation of the Ligation Products
.........Principles of DNA Transformation
.........Vectors
.........Blue/White Selection
.........TA Vector Discussion
.........Antibiotic Selection, etc.
Start Minipreps from Plates.
.........Sterile Techniques
.........Redo transformations and/or PCR

Week 7/8
Minipreps on all Picked Colonies
.........ETOH ppt
.........Speed Vac Minipreps - Resuspend in TE
.........Electrophoresis "uncut" DNA

LAB Take-home Test#1 Given Out (Due in 1 week)

Week 9/10
Review Principles of Restriction Digestion
.........Do Eco RI Digestion on Miniprep DNA (Detect Inserts)
.........Run the digestion products out. Size of Inserts?
.........Sequencing Principles

Week 11/12
Sequencing Principles-Radiation Safety
.........Fetamolar Kit - Setup
.........Sequencing Rxn.
.........Run Sequence Gel - Load

Week 13-15
Principle of Reading a Sequence Autorad
.........Read Gel
.........Darkroom Work
Internet for Biologist
.........Genebank-Submit Sequences to Blast
.........Analysis of the your group’s data

LAB Take-home Test#2 Given Out (Due in 1 week)

LINKS for Molecular Biology Class
NIH Blast
What is Blast?
Blast FAQs
National Center for Biotechnology Information
National Library of Medicine
National Institutes of Health
Virtual Genome Center
Entrez
PubMed
The Dictyostelium Virtual Library
National Science Foundation
Dr. Bush's Latest Grant Abstract: NSF #9734093

*****Special Programs At UALR*****

The Molecular Biotechnology Program
The new degree emphasis in molecular biotechnology is designed for students that want to pursue a research career in molecular biotechnology at the bachelor’s level. This concentration will prepare students for technical occupations in basic and applied molecular biotechnology research.
www.ualr.edu/~biology/biotech.html

Graduate Program in Biotech -Applied Science Department at UALR


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