Sauer Lab
Lab members on OWW
Former lab members on OWW
Protocols
Here we will post protocols that people in the lab generally use. Many of these will be lab-specific protocols, but if you think the protocol you are posting is more generally applicable, please link it to the general Protocols page. Also, if you typically do a variation of the protocol that is posted, please list it on the same page as a separate, alternative protocol. If possible, also try to note why what you do is different.
PLEASE NOTE: Blindly following a protocol is not a good substitute for actually talking to someone who has done the experiment. When posting protocols, please also include the name(s) of people in the lab who will be good to talk to about the protocol.
Sauer:In vitro degradation of GFP-ssrA by ClpXP
Sauer:Expression/purification of 35S-Met proteins
Sauer:In vitro degradation of radiolabeled proteins by ClpXP
Sauer:Quantitative Western blot
Sauer:Site-directed mutagenesis
Sauer:Competent cell preparation
Sauer:P1vir phage transduction
Sauer:In vivo tmRNA tagging assays
Equipment (Sauer lab)
Please list here the equipment we have in our lab and the appropriate contact person for that piece of equipment. If it is something a lot of people use, also list other people that may be able to help out if there are any questions.
Thermocyclers (PCR machines)
Located in room 565
Sign-up sheets for both machines are located nearby. Please use them, even if no one else is signed-up/using the machine. This lets people know where to put your reactions when they are finished if they need to use the machine, or who to talk to if they want to know when you're going to be finished.
No one is officially in charge of these. If you need help, ask Kathleen, Sean, Andreas, Dan, Chris, Eyal, Brent, Joe, or Greg.
- DNA engine
- Status quo set up has two independent blocks that can each hold up to 30 ~0.5 mL tubes
- We also have a gradient block (located on the shelf above the machine). This block holds one microtiter plate (96 well) or up to 96 0.2 mL tubes. You can set up a temperature gradient with this block to optimize your annealing temperature. To use this block, you need to remove the block with the two independent units and replace it with this one (turn the machine off first and ask someone/RTFM). Note that this means that no one else will be able to use the machine when you are using it, so please sign-up accordingly.
- Both blocks have a "hot top", so there is no need to use oil on top of your reaction.
- Perkin Elmer Cetus DNA thermal cycler 480
- affectionately referred to as "Bessie"
- This machine is an "old school" peltier thermal cycler. It holds up to 48 ~0.5 mL tubes. No hot top on this one, so be sure to use mineral oil on top of your reaction to prevent evaporation
Equipment (other labs)
Please list here equipment that we use in other labs along with a contact person in our lab and a contact person in the other lab. Tips on how reserving/using the equipment works would also be helpful i.e. do we need training on the equipment?.
