Fall Semester 2005 Activities subject to the uncertainties of science. Deadlines are firm. Updated Oct. 13, 2005 NOTE: Readings and handouts are available by direct link or via e-res, the electronic reserve system of the CONSORT libraries. |
Sept. 1 | Course introduction; pipettor verification; quantification of nucleic acids by spectrophotometry; intro. to degenerate primer design | Morrison et al. (1995) Morrison et al. (1998) Iwata and Stegeman (2000) Wilkie and Simon (1991) |
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Sept. 8 | Total RNA Isolation, quantitation. | RNA isolation protocol | |
Sept. 15 | RNA gels, photography; Figure preparation; Workshop: Primer design. |
Wilkie and Simon (1991) | Homework: primer sequences |
Sept. 22 (Meet in Computer Lab, FSH 009) |
Computer Workshop: Bioinformatics BLAST searches, sequence alignment, phylogenetic trees. | Handout: DNA and Protein Sequence analysis. |
Lab Report 1: RNA Isolation Also: Notebook check. |
Sept. 29 | RT-PCR for CYP1A cDNA fragments | RT-PCR Protocols | |
Oct. 6 | Agarose gel analysis of PCR products | ||
Oct. 13 | Purification of RT-PCR products from gels; ligation of products into plasmid cloning vector. | Gene Clean protocol Ligation protocol |
Lab Report 2: Sequence analysis exercise. |
Oct. 20 (and 21) | Transformation of competent E. coli cells. | ||
Oct. (26 and) 27 | Plasmid minipreps; restriction enzyme digestion. | Miniprep protocol Restriction ezyme protocol |
Notebook Check by request. |
Nov. 3 | Gel analysis of miniprep digests. Writing workshop: Guided peer review previous lab report. |
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Nov. 10 | Workshop: Intro to Real-Time PCR; |
Applied Biosytems Real-Time PCR Tutorial. | Lab Report 3: RT-PCR and cloning |
Nov. 17 | Real-Time PCR Reactions on ABI 7500 | Real-Time PCR Protocol | |
Nov. 24 | Thanksgiving Break |
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Dec. 1 | Real-Time PCR Data Analysis | ABI 7500 Tutorial | Lab Report 4: Analysis of YOUR sequence |
Dec. 8 | Course catch-up and wrap-up: discussion of results and check-out. | ||
Dec. 19 (Monday) | FINAL REPORT AND NOTEBOOK ARE DUE: 11:30 AM (corresponds to the end of the schedule exam time). | FINAL REPORT DUE 11:30 AM |
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Last Modified Oct. 13, 2005