Chem 125L
Laboratory Techniques in Biochemistry
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Chem 125L |
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Dr. Kalju Kahn
Office: PSB-N 2623
Office hours: Tue 11:30-12:30, "Open Door" policy other times
Phone: 893-6157
E-mail: kalju@chem.ucsb.edu
Website: http://www.chem.ucsb.edu/~kalju
Section 1: Paul Zakrevsky pzakrevsky@chem.ucsb.edu Office hours: TBD Phone: x5302 Section 2: Javin Oza oza@lifesci.ucsb.edu Office hours: TBD Phone x2677 Section 3: CLOSED
Chem 125L is an undergraduate laboratory course where students will carry out a project similar to that performed in many contemporary biochemistry and molecular biology laboratories. The project involves changing specific amino acids in a protein by site-directed mutagenesis of the corresponding plasmid DNA, expressing the mutated protein in the bacterial cells, purifying the overexpressed protein by means of column chromatography, and characterizing properties of the mutant protein.
Lecture: Mon 9:00-–9:50 AM in Phelps 1160
Lab section 1: Tue 2:00-5:50; Thu 2:00-5:50;
Lab section 2: Tue 6:00-9:50; Thu 6:00-9:50;
Lab section 3: CLOSED
Lab sections are in PSB-N 2619
| Syllabus | General information about the course. | |
| Upload | Submit your proposal as a PDF file | Link |
| Guidelines | Write-up for Component I: Due Feb 14 | |
| Guidelines | Write-up for Component II: Due Mar 6 | |
| Guidelines | Write-up for Component III: Mar 15 | |
| Guidelines | Research Presentation: | |
| Guidelines | Final Exam: |
Students in the class do not have to purchase the laboratory manual. The theory manual and chapters of the operations manual can be downloaded here in the PDF format. Please note that you can follow hyperlinks that are in the PDF files by clicking on the link.
| Experiments | Download Adobe Acrobat Here | Acrobat |
| General: | Theory Manual | |
| Background | Structure and Function of DNA | |
| Background | DNA Technologies | |
| Planning | Operations Manual, Pre-proposal work | |
| Planning | Sample NSF grant proposal | |
| Comp I | Operations Manual, Component I | |
| Comp II | Operations Manual, Component II | |
| Comp III | Operations Manual, Component III |
Many experiments in this course involve use of materials or equipment for which manufacturers have provided nicely illustrated operations manuals. Normally, you want to obtain the most up-to-date versions of these manuals from the manufacturer's website. Copies of some of these documents are provided here for your convenience.
| Literature | Required or optional reading in PDF | Acrobat |
| Comp. 0 | How to Write a Grant Application | |
| Comp. 1 | Stratagene QuikChange© Kit: Manual | |
| Comp. 1 | Stratagene XL1 Competent Cells: Manual | |
| Comp. 1 | Stratagene BL21 Competent Cells: Manual | |
| Comp. 1 | QIAprep Miniprep Handbook | |
| Comp. 2 | BioRad Bradford Assay Manual | |
| Comp. 2 | BioRad Chromatography Guide | |
| Comp. 2 | BioRad Hydroxyapatite Guide | |
| Comp. 2 | BioRad EconoPump Manual | |
| Comp. 2 | BioRad Fraction Collector Manual | |
| Comp. 2 | Ni-NTA Spin Column Manual | |
| Comp. 2 | BioRad MiniProtean 3 Manual | |
| Comp. 2 | Stirred Ultrafiltration Cell Manual | |
| Comp. 2 | Ultrafiltration Disks Manual |
If time permits, I will post short summaries of what we discussed in the class.
| Week 1 | Experiments in Teaching and Research Settings. DNA biochemistry. | |
| Week 2 | Bacterial DNA metabolism; DNA methylation. | |
| Week 4 | Protein expression, gene regulation, and cell lysis. |
Camera images and student files on the C:\Students\ disk on the computer are in: TR2 TR6 WF2
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