Welcome to the Macara lab!

Important information for new (and old) members of the research group:

We have tried to organize the Macara laboratory to run EFFICIENTLY, so that you spend as little time on mundane tasks such as making buffers, and as much time as possible doing actual research.

But - The laboratory does not run itself!

Without your help it will grind to a halt.

Keeping the lab running smoothly requires TEAM EFFORT.

This means being a good LAB CITIZEN!

This means that EVERYONE – NEW LAB MEMBERS AND OLD – play important roles in maintaining lab functions.

This is important for safety reasons as well as efficiency.

There are some basic rules that EVERYONE needs to follow, EVERY DAY:

GENERAL STUFF:

CLEAN UP AFTER YOURSELF.  Especially clean up the BALANCES themselves, and the area around the balances.  If you do not clean the balances, they will corrode, and become inaccurate.  Use a damp tissue to clean.
Rinse SPATULAS and STIR BARS in d. water, dry with a clean Kimwipe, and place them back in the drawer.
If you use the last of any REAGENT, write it up immediately on the BOARD, so a replacement can be ordered – or submit the request on QUARTZY.  Please include CATALOG #, COMPANY, PRICE, and QUANTITY.
If there is little left of the reagent you are using, WRITE IT UP on the board or enter into QUARTZY! –as above.  Do NOT put empty tubes back in the freezer.
Discard outdated, contaminated or incorrect materials – do not let them accumulate in the freezers or refrigerators.
If you take a new tube of enzyme out of the backup boxes, submit on QUARTZY or write it up on the board so it can be replaced.
If you sign for new reagents, etc, when they are delivered to the lab, be sure to tell Amanda.  Only put stuff away if you know where it goes! – do NOT just stuff restriction enzymes etc in the -20 freezer.  Put them in the proper box, enter the information in QUARTZY.  Put the invoice into the folder next to Amanda’s desk.
Keep the WATER BATHS full! – if they dry out, the heating element will be damaged, and a replacement water bath will cost up to $1,000!
Make certain that all tubes are balanced before putting them into a centrifuge.
If bottles leak when CENTRIFUGING, remove and rinse the rotor, and drain upside down on the equipment table near the centrifuge.
If anything is running low (buffers, reagents, nitrocellulose sheets, competent bacteria, etc, etc) write it up on the WHITE BOARD – better safe than sorry!!
Do not put spatulas, stir bars, centrifuge bottle caps, or Tupperware on the dish cart.  Wash these items yourself and PUT THEM AWAY WHERE THEY BELONG.
For permanent lab members:  One lab member a week (weekly schedule by dish cart) is responsible for LB, sterile water, 1x TAE, sterile bottles and 1x reservoir buffer.  Check the schedule!
NEVER leave stuff that needs to be autoclaved out on the bench overnight – it will become contaminated, and will contain toxins that might survive autoclaving.
If you don't know how to use something – a centrifuge, microscope, pH meter – whatever – ASK SOMEONE TO SHOW YOU HOW TO USE IT.
If there is a mess in the lab and it is yours, clean it up. If it cannot be determined who made 
the mess, take your turn being a good citizen and clean up the mess even though you had nothing to do with it. “Always leave the camping area cleaner when you depart than it was when you arrived.”
 Turn OFF equipment at the end of day.  Last one in the lab? – BEFORE YOU LEAVE, CLEAN UP, MAKE SURE EVERYTHING HAS BEEN PUT AWAY (CHECK WATER BATHS), TURN OFF THE LIGHTS, TURN ON THE UV LAMP IN THE HOODS.

MOLECULAR BIOLOGY:

When using enzymes in the cooler boxes, NEVER EVER put a pipet tip twice into a tube – cross –contamination would be disastrous for you and everyone else!
ENZYME BOXES – last out of the freezer, first back in the freezer!  And clean off the ice before returning them to the freezer.
NEVER use enzymes in the BACK-UP BOXES unless the tube in the enzyme box is empty – then throw out the empty tube, move the new one from the back-up box to the enzyme box, and submit a replacement order to QUARTZY.
After running gels – Rinse out the gel boxes and put them away.  Clean the gel trays and put them away in the drawer.
If you make a new vector, or receive reagents from another lab, LOG IN using the books in the computer office, and enter it into Quartzy.  Store at least one aliquot of bacterial stocks in the common boxes in the –80 freezer, and LOG IT IN (with date, origin, etc).
When ordering oligos from IDT – check with others in the lab to pool orders if possible – oligos are cheap but shipping is VERY expensive.  Or us the MBCB CORE, which has free shipping
DNA sequencing – use a fresh 3ml overnight culture; use the Qiagen miniprep kit; measure the DNA concentration; add 1 ul of the 10 uM primer stock solution (NOT the 100 um stocks used for PCR!!).  These are stored in the white –20 freezer door.

CELL CULTURE:

CLEAN UP AFTER YOURSELF!  Wipe down the hood with 70% ethanol.
Make sure the tray in the base of the incubator has water in it.  If it dries out your cells will die!  The autoclaved water is in a large conical flask on the table just outside the room.  If you use the last of this water, refill with MilliQ water and set it out to be autoclaved!
Turn off the LAMP on the microscope when you have finished with it.
WRITE UP ON THE BOARD if aliquots of P/S, serum, bottles of trypsin, DMEM, or transfection reagents are running low, and tell Amanda.
If you use up the last Pipetman tips, put the box out to be refilled (under table outside Ian’s office), and REPLACE box from the storage cupboard.  Also replenish ethanol, pipets, dishes, etc as they are used up.

FINDING THINGS:

We use QUARTZY as our web-based lab inventory and for ordering. It also contains PDFs of a variety of useful protocols.

IMPORTANT - Log all new antibodies, plasmids, enzymes, etc into this inventory when you receive them or make them and include the location! – the inventory is only as useful as people make it (think Wikipedia!).       

OTHER STUFF:

Lab Notebooks and Labeling:

It is required that you keep a “hardcopy” notebook. We use hardbound “essay book” for this. When you have electronic data you can refer to it (with filenames and computer locations) in this.

Date every entry in your notebook, and include a brief statement of purpose. Provide a protocol/description.  Include all details of each experiment -  mistakes, failures, changes to the protocol, as well as calculations, dilutions etc.  It is often helpful to check off reagents as you add them to tubes to avoid screwing up the experiment if you are interrupted.

Tape in copies of your data - blots, etc; and make notes of filenames and exactly where online images etc are being stored so in 2 years time you can quickly find them again.
Lab notebooks are the property of the lab.

Do NOT accumulate weeks' worth of notes on scrap paper - update your lab book daily.

Figures:

When you present me with plotted data please Include full details of what the data represents. Label figures with temperature, pH, concentrations of protein, lipid, and detergent etc.  I should be able to look at your data 5 years from now and know what it is.
Try to plot data at publication quality. This means using font size and boldness that is clearly legible and using pen sizes for axes that are not spiderweb-like. And so forth. I would like to be impressed not only with your beautiful data, but how beautifully you present the data. It helps your reputation. Fill the page with the plot (don’t make a 2” X 2” plot on a 8.5” X 11” piece of paper).

Remember that many journals now REQUIRE entire blots from which relevant data has been extracted for figures to be made available to readers - make sure you RETAIN all original blots and other data.