Biology 22 Laboratory Report I 1. Bacterial Transformation 2. Plasmid Isolation 3. RFLP Analysis

Also you can refer to your syllabus which has a written description of the lab report.

Format of Laboratory Report Typed  Title Page  Provide a descriptive title  Your name  Must include group numbers (not individual lab partners) for each experiment  Results  All tables have titles and numbers above table  All figures have legends and numbers below figure  Discussion: 2.5 to 3 pages double spaced  If discussion is more or less than the 2.5 to 3 pages, then points will be deducted.

Results Transformation results Transformation efficiency Photograph of isolated plasmid Photograph of RFLP Standard Curve Table that summarizes the size of the fragments of each lane Individual Maps

Results: Transformation Table: results of 3 agar plates: number and color of colonies Transformation efficiency

Results: plasmid isolation  The photograph of the plasmid isolation.  You should include a numbered legend to this photograph. The legend includes: what samples are present on the gel, their “lane” number, and the type and percent gel.  Note: The gels were a 0.8% agarose

Results  A photograph of the RFLP analysis with figure legend and number

Results  Standard curve Distance migrated (cm) DNA bp

Results  A table that shows the bp versus distance migrated of your unknown fragments  A separate table that shows the the bp versus distance migrated of your ladder (known) fragments

Examples of MAP Results  These are just examples! Total Size: 7000 bp Lane 3 Ecor I Lane 4 Total size 7050 bp Mst II

Number of Maps Lane 3 Lane 4 Lane 5 Map 1 Map 2 Map 3

Discussion On the next couple slides are some questions to help guide you through the discussion of lab report 1. They are meant to help you think about your data, but please do not hand a question/answer format to your lab report. These questions are for you to read independently but they are concepts that you should address in the report. However, I also encourage you to think of other concepts that you might want to address.

Put together a logical story based on your data and be creative. Lastly, try to enjoy yourself as you put the pieces together

Concepts/question to address in your discussion: Transformation 1. What did the results from the transformation show about how the plasmid altered phenotype? 2. What reason(s) would you expect for the fact that transformation efficiency is often times quite low? 3. What is the mechanism for the transformation? 4. Why are ampicillin and X-gal included on the agar plates? 5. What is the purpose for including control plates.

Discussion Plasmid Isolation 6. How was it possible to separate cellular proteins from nucleic acids? 7. Why did the photograph show more than one band for the plasmid isolates? 8. How did you know if RNA was present and how could it be removed?

Discussion RFLP 9. How do restriction enzymes create unique patterns? 10. Which restriction enzymes are unique enzymes? 11. Which enzyme might be used for a cloning experiment to “insert” a piece of DNA into the plasmid? 12. What are some other possible applications for the RFLP technology other than for simply mapping a plasmid?

Discussion  Do NOT simply list the questions and present the “discussion” as a “Q and A” section.  The questions are meant to help you address important concepts of each experiment.

Discussion   Everyone must do their own work, please do not copy from each other nor plagiarize from the internet or book. If evidence exists of either of these practices, credit will not be given for either your report or your colleague’s.

The Do Not’s Do NOT copy out of the lab manual (or any other source). Do NOT make the focus of the discussion a list of errors. Do NOT simply state the data cannot be interpreted because of errors—this is just not true! An effort has been made to see that you have access to photos and data that allow you to meaningfully interpret your experiments. If you had a specific problem with an experiment this should be mentioned in your report but then be sure to discuss the data made available to you. Do Not

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