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
Sauer:In vitro transcription with T7 RNA polymerase
Sauer:Phenol/chloroform extraction
Sauer:bis-Tris SDS-PAGE, the very best
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.
Sauer:Thermocyclers (PCR machines)
Sauer:Analytical ultracentrifuge
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?.
Other lab stuff
FAQs
Q: Can we get this in the stock room?
A: Check out MIT ECAT, VWR page (certificates required). There is a listing of items carried in the stock room (link is on the left side of the page).
Q: What if it's not in the stock room, but VWR carries it?
A: Order the item through MIT ECAT. You will need an account number (get this from Brenda) and you will need to know the G/L account number to use (if you don't already have a list of these, ask Kathleen). You can also use ECAT to order from other companies, like Office Depot and Airgas.
Q: How do I submit samples for DNA sequencing?
A: Check out the MIT biopolymers site. The facility also does mass spectrometry, peptide synthesis, peptide sequencing, and a few other things. You can download all of the forms you need from the site and there are instructions on how to retrieve your data. You will need an account number (talk to Brenda) to submit samples. Talk to someone in the lab about our login and password for the DNA sequencing site. Whenever sending things to biopolymers, be sure to give Brenda a copy of your submission sheets. This helps with questions about charges later on. Also, for DNA sequencing, keep a copy for yourself. Data is posted as 1, 2, 3,..., so it is important for you to remember which sample corresponds to each number.
Q: How do I order things that you can't use a credit card for (like needles, syringes, ethanol, radioactivity)?
A: If these things can be ordered through VWR, MIT ECAT is the easiest way to get them. If you can get them in the stock room (needles, syringes), use an MIT requisition form. These can be found in each room in the bin under the packing slip bin. For other things (like radioactivity), you need to create a requisition through MIT SAPweb. After creating the requisition, check back on the site later to get your purchase order number. Then, use the purchase order number when you call the company to place the order. If you are using this for radioactivity, ask someone who has done it before for help because certain non-obvious things need to be included on the request form.
Miscellaneous links
- Login required
- Certificate required
- Purchase orders, etc. Certificate required.