Week |
Date |
Discussion Topic/Lab |
Notes/Papers |
Figures |
Labs |
1 |
9/8 |
Discussions: Course Introduction
Why is transcription important for biology and human diseases?
Review of of the general process of transcription and reporter genes
Lab: Learn to use pipetors |
L1-Transcription
Editorial |
L1F
|
Lab-0 Introduction
Lab-1 Pipeting
|
2 |
9/15 |
Discussions: Why do we work in yeast? Review of Yeast Biology
Genetic analysis of transcriptional regulatory sites
Genetic
screens for activator and repressor proteins
Lab: Sterile Technique, Plate mutagenized yeast |
L2-Yeast
|
L2F
|
Lab-2 Plating |
3 |
9/22 |
Discussions: Xie et al.
Activity of MSEs from mid-sporulation promoters
Identification of transcription factors that function at MSEs
Methods - lacZ reporter assays
Lab: Streak colonies to screen for silencing and repression assays
|
L3-MSE
Xie et al. |
L3F
|
Lab-3 Streaking Colonies |
4 |
9/29 |
Discussions: Xie et al. cont.
Methods - lacZ reporter assays, Eukaryotic chromatin structure
Silencing vs repression, Sir2 vs Hst1; PCR Mutagenesis
Lab: Perform filter lifts and lacZ assays
Analyze repression, silencing - pick mutants to sequence
|
L4a-Silencing
L4b-Minipreps
Mead et al
McCord et al.
Sir2-Aging
Sir2 Review |
L4F |
Lab-4 LacZ Assays |
5 |
10/6 |
Discussions: Xie et al and Mead et al.
Eukaryotic chromatin structure
Silencing vs repression, Sir2 vs Hst1; PCR Mutagenesis
Lab: Plasmid Minipreps, Set up ON yeast cultures |
Digets, agarose gel |
L5F
|
Lab-5a Plasmid Minipreps
Lab-5b Set up ONs
|
6 |
10/13 |
Discussions: Review of Screen data; Sequence analysis, 4-Peaks, FinchTV, Make a contig
Identify the mutants
Lab: Restriction Digests; Spot assays
|
|
L6F
|
Lab-6a Restiction Digests
Lab-6b Spotting Assays
|
7 |
10/20 |
Discussions: Sequence analysis, 4-Peaks, FinchTV, Make a contig
Identify the mutants
Lab: Agarose gel electrophoresis
|
Cn3D Instructions |
L7F
|
Lab-7 Agarose gels
|
8 |
10/27 |
Discussion: Review of Sir2 structure; Structure modeling using CN3D or RasMol Results, problems and interpretations
Lab: Analyze sequences, perform BLAST, map mutations on the structure
|
|
L8F
|
Lab-8 DNA Sequence Analysis
|
9 |
11/3 |
Discussion: Review of Sir2 structure; Structure modeling using CN3D or RasMol Results, problems and interpretations
Lab: Analyze sequences, perform BLAST, map mutations on the structure |
|
L9F
|
Lab-9 DNA Sequence and Protein Structure Analysis |
10 |
11/10 |
Discussions: Presentation of data, Interpretation of the results
Powerpoint files are due by 10:00 AM on 11/10
Lab report with discussion of the data is due by 1:30 PM |
Presentation Outline |
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