"If you want truly to understand something, try to change it"
This page is the PRELIMINARY homepage of the course titled "840:153g". Current version: Fall 2008! The page and course contents are still under development - please check back often!
During this course, you will be introduced to basic techniques for cloning and recombining DNA. The principles of gene cloning, DNA manipulation and analysis will be addressed from a practical view. This lab requires designing your own experiments, self experimentation, record keeping, and discussion and reporting.
Upon completion of the course, you will
- understand how gene sequences are translated into protein in different organisms
- conceptually understand how to manipulate DNA and create recombinant proteins or modify protein expression
- be able to identify, analyze, and functionally recombine DNA molecules
By taking this course, you will learn how to isolate, analyze, and clone DNA. You will have a broad overview over the most common procedures used in a molecular biology laboratory. You will be able to design and carry out simple experiments, document and explain your results. You will have demonstrated that you developed skills in finding and using genomic information stored in public repositories, analyze and summarize your findings, and logically organize and present your data.
You will work in teams of four (yes that's right: 4) throughout the course. Each team will design a new biological part or device, assemble it, prove it, and store it in the registry of biological standard parts. Your team may use all parts stored in the registries 2007 DNA distribution kit. The kit is available to you in form of DNA. Therefore, see these instructions how to use the kit.
Like in the International Genetically Engineered Machine (iGEM) competition, you will work in teams to develop a new biological part, module, or device. Our teams are smaller, but I expect each team to develop something new - something that does not yet exist in the repository of biological standard parts. This is a very challenging task for the few sessions we have and not necessarily something all teams will be able to complete. Therefore, very good planning is required. The better your team plans the entire project from the beginning, the better your chances are to complete your project. Since good planning is very important, it will contribute 20% to your grade. Your team has to carefully describe your project on your project homepage and give a presentation during the third week of class.
It is up to each team to make a decision which new part to make. However, teams should carefully study the parts list to find something that's not already in there. New parts can be as simple as a new protein coding device or regulatory region, a combination of existing parts, or something else you would like to do. However, we cannot synthesize DNA because that is too costly. Therefore, you have to work either with existing parts (from the 2007 repository) or use DNA that you can amplify by PCR from readily available resources (for example food plants). In addition to the particular part, module, or device, your team wants to produce, each team needs to make sure that parts are biobricks compatible. This means your part must eventually be modified by site-directed mutagenesis to substitute for conflicting restriction sites (e.g. those used in the biobricks standard assembly). Finally, you (as a team) have to provide evidence that your part, module, or device is functioning in the way you predicted. Each team presents its results in another presentation after thanksgiving week. This presentation is again worth 20% of the grade.
How to Start
1) Select your part, moduel, or device
Your first team task is to identify a new part, module, or device your team wants to produce. Browse the parts in the registry to find out what other people did, what can be done, what is available to use. Then decide what your team wants to do. Be curious and select something interesting. Don't be afraid to select something that you eventually might not be able to complete during the semester. The only limit you have to consider is the availability of a source for cloning (e.g. a tissue sample).
2) Describe your project on the team homepage
You have to describe your project on the team homepage. The description should be very concise and contain enough details to understand the project in all its details. That includes:
- desrciption of the product (part, module, device)
- sources for cloning it (e.g. tissue, parts used)
- steps required to make it
- proof of function
Developing your ideas and figuring out what is involved is a complex task that involves all team members. You have to sit together and communicate your ideas. Brainstorm ideas, make plans, assign work pieces to everyone, find information, discuss what you found, revise your concepts, look up protocols and describe your approach on your homepage. Familiarize yourself (everyone) with OpenWetWare and how to edit pages. Do not forget to apply for an account after (or during) the first class session. You will need it immediately. The sooner you start writing (even rough drafts, lists of ideas) the easier it will be for your team to keep track of ideas and find relevant information. And the more you write the better your grade will be. WRITE IN EVERY SESSION! The completeness and clarity of your strategy will be assessed and graded (20%) by the end of the third week. The approach must be written down in form of a research proposal on your project website and approved by the instructor before lab work can commence.
You almost certainly will identify a couple of gaps in your knowledge that have to be addressed in order
to complete or even start your project. Questions you cannot solve on your own (until your proposal is due at the end of week 3) should be listed at the end of your experimental strategy under the "Need to Know" section. Ask yourself (and your team member): What information (about the
part and the experimental procedures) do you need to know in order to produce and verify it? Prepare a list of
issues you need to address! Keep in mind that the WIKI pages can be easily edited. So start writing as soon as possible. You can revise and modify it later.
Please be aware that the first 2-3 weeks may indeed be spend developing your experimental plan rather than working in the lab. If
you wish, you can consider this as the combined lecture part of the course.
Attendance at ALL laboratories is mandatory. Only religious observance, a death in the family, or serious illness/injury and a doctor\\\\\\\'s note are valid excuses. One unexcused absence from lab will result in failure of the course. Lecture and lab sessions are always combined. There will be no formal lectures but short lecturings on specific topics may be given occasionally and if needed.
Your team will need to develop a website at OWW in which you describe your project (in form of a proposal). The first 10% of the grade will be given for selecting a gene, developing an experimental outline, and completing the project description. This is due by the end of week two.
A large and very important part of the class grade will be based on the writing notebooks. A lab notebook is required that will be graded based on completeness, accuracy, and content. It should contain all experimental procedures carried out during each lab session (incl. observations, calculations, remarks, etc.), as well as description and discussion of results. Based on the results obtained, it should also contain a brief description of the steps you plan and materials you need for the next session. The notebook has to be written at the day of the lab - during the lab! Depending on these notes, an online journal must be kept. This journal has to be updated before the next lab session. No late entries (even 1 day) will be considered. Since each entry in the online journal receives a time stamp and the name of user who writes it, it very easy for me to monitor everyones contribution. Notebook keeping is considered an essential part of the work and therefore should be done during the lab sessions. There will often be lag times while experiments run. Use them for writing your lab notebook and updating the online journal (the computer room is right next door).
Finally, I expect each group to clone their fusion gene. If a group does not achieve it, the problems must be explained thoroughly in the online notebook and evidence must be provided from the lab notebook that sufficient efforts have been undertaken to clone the gene. After all, this is real science (none of the genes you will work with have ever been cloned) and there is no guarantee that any group will succeed (though chances are high). Therefore, keeping a good notebook is essential. The notebook will account for 40% of the grade (20% for online journal, 20% for the written notebook) of the grade and both partners receive the same grade for the notebook, weighed by a peer assessment. Both partners, however, should regularly switch responsibility to write notes.
I will assign 30% of the grade based on everybody's individual contribution. Here, I will consider contributions during class sessions, including preparation for class, discussion of experiments and results, timeliness, and teamwork. The remaining 20% will be given for the project presentation. One member of each team will give a midterm presentation (around mid October), the other member will give a final presentation (the week before finals). Both presentations should give a 10-minute overview of what has been done - and achieved - in the project. It will be essential to provide experimental data, describe and explain them carefully, and discuss them with classmates.
Final Grade Composition:
- 10% project description and experimental plan (due week 2)
- 20% lab notebook
- 20% online journal
- 30% individual contribution (preparation, performance, knowledge)
- 20% project presentation
Once again: the three first grades will be team grades for which both team mates will receive identical grades. The last two grades will be individual grades. There are no make-ups possible !!!
Getting Started with OWW
To get started with your online journal, you need to apply for an account at OWW (which is free). Please write down your username and password from your confirmation email. After you obtained your account details, please go to the Project Page and start developing your project. There is help available on how to use this WIKI in the left-handed navigation pane and in the top menu bar.