The experiments described in these materials are potentially hazardous and require a high level of safety training, special facilities and equipment, and supervision by appropriate individuals. You bear the sole responsibility, liability, and risk for the implementation of such safety procedures and measures. MIT shall have no responsibility, liability, or risk for the content or implementation of any of the material presented.
The following is an index of laboratory protocols, downloadable in pdf form, that are used by students in this course.
|AN12 Defined Medium||(PDF)|
|Beta Galactosidase Assays||(PDF)|
|Biotin Detection in Electrophoretic Gels||(PDF)|
|Bradford Assay for Determining Protein Concentration||(PDF)|
|Conjugal Transfer with B264-1||(PDF)|
|Defined Medium for Rhodococcus SPP I24 and KY1||(PDF)|
|Electrocompetent Corynebacterium and Rhodococcus B264-1||(PDF)|
|Electrocompetent E. coli||(PDF)|
|Electrocompetent Rhodococcus AN12||(PDF)|
|Electrocompetent Rhodococcus I24 and KY1||(PDF)|
|Electrocompetent Rhodococcus SQ1||(PDF)|
|Ethanol Precipitation of Nucleic Acids in Aqueous Solution||(PDF)|
|Gel Loading Dye||(PDF)|
|Gel Running Buffers||(PDF)|
|Genomic DNA Preparation from E coli Cells||(PDF)|
|Genomic DNA Mini-Prep||(PDF)|
|Genomic DNA Preparation from Yeast||(PDF)|
|Maxiprep of Plasmids from E.Coli using Wizard Maxiprep Kit||(PDF)|
|Partial Digests of Plasmid DNA||(PDF)|
|PCR Amplification from GC-Rich Templates||(PDF)|
|Phenol Extraction (for DNA Purification)||(PDF)|
|Primer Design for PCR Amplification and Sequencing||(PDF)|
|Quantitating Nucleic Acids||(PDF)|
|Southern Blotting with Roche DIG Labeling System||(PDF)|
Guidelines and Recommendations for Lab Notebooks
The notebook is the key to good experimental practice. It should be a complete record of all the experiments as they were actually performed. A good notebook will enable someone to reconstruct, long after the fact, exactly what was done and why.
Laboratory notebooks should be bound with numbered pages. In the notebook you should write down, as you go along, what was actually done. Experimental results, tables, graphs, and photographs should be written directly in your notebook or taped to the pages for a permanent record.
Your notebook should minimally include the following information:
- Table of contents
Leave extra pages at the beginning of the notebook for a table of contents, which you'll add as you go along. The table of contents should have a short title of each experiment, numbered and dated.
- A short summary of the reasoning for the experiment
Often a single sentence will suffice. You may want to include a summary of the method you will use.
- All the relevant information about the preparation of the experiment
Much of this can be accomplished by suitable cross references to previous experiments. The first time a solution is introduced, its origin should be noted. If a component or method is used for the first time, then it should be described in detail. The same should apply to bacterial strains, plasmids, etc.
- The actual series of events as performed
Especially important here are the actual times of incubation (as opposed to what was planned), any changes in procedure, any unusual observations made, etc. Writing down what actually happened can make it possible to interpret an experiment even though many mistakes were made.
- The results
Include raw data in the form in which it was collected. For example, record the actual number of colonies found, as well as the cell concentration calculated from this number. A brief summary of the results and interpretation will also be helpful when reviewing the data at a later date.
The main point is that it is unwise to rely solely on memory. The staff urges you to write directly into your notebooks and not to waste time copying things down later.
After acquiring some experience, you may want to know something about an early experiment which did not seem important at the time. If you are in the habit of writing things as you go along, the chance of being able to learn from the early experiments is much greater.