840:153g:Materials

Course materials

This lab course is designed to provide a real life experience of work in a genetic engineering lab. Therefore, you will have to prepare project specific media and solutions yourself. Most of the chemicals you need will be available, but not all solutions and buffers will not be ready for you. You have to make them yourself if you are using special protocols (for example plant specific DNA or RNA extraction procedures). If you need something that you cannot find, ask the instructor or Larry in the biology stock room. You need to check if you have all your buffers and solutions available one session before you start your planned experiments! Please think ahead and let me know of any special needs you may have. If you don't plan for your next session, your work will very likely be delayed (which will affect your grade). I cannot provide special solutions or buffers, or sterile media or Agar plates with appropriate antibiotics ad hoc. And nobody will prepare them for you! Since each project will progress in its own pace it is impossible for me know in advance what you might need. Therefore, ask if you are not sure. But do not assume I can step in if you forgot to make something. I cannot. I need you to plan ahead to prevent unnecessary delays. Your careful planning is part of the course and will be evaluated. You also will learn to prepare basic solutions by yourself. This will become very important in case you end up working in a lab, either on the job or in grad school. Therefore, it is fully your own responsibility to plan ahead and prepare what you need for the next lab. Think ahead and read the next steps in the protocol. Otherwise you will loose important lab time and this will effect your performance (and your grade). I would strongly suggest to put a "needed items" list into your notebook so that you know what to you need to take care of before the next lab session. Please talk to me frequently and as early as possible about your needs so that I can help you find the materials (or alternative ways).

Some material we have available for you

• we use FastDigest Enzymes from Fermentas
• we use 2x PCR Master Mix (Choice Tag) from Denville
• we use 1x TBE buffer to run Agarose gels
• we use either of the following two DNA molecular markers from Fermentas (please use both sparingly - they are very expensive):
  • 50 bp ladder:
  • 1 kb ladder:
• we use antibiotics (Amp, KAN, Tet, ...) in 1000x stocks. That means, you must use 1 mL per L of medium!
• X-GAL and IPTG stocks are also 1000x concentrated

Useful protocols (from OWW)

DNA protocols

PCR techniques

E. coli protocols

Plant DNA isolation

Plasmid miniprep