Rules

Note: These rules are very strict; if you fail to comply, you will be used for experiments.

1. Safety: Always wear your lab coat in TC room when working with cells.

2. Space: Everyone has individual refrigerator, shelf and drawer space for their reagents, including media bottles, pipettes and plasticware. Keep common space clean. If you're running low on space, just ask Jane.

3. Community Supplies : When you're the last to use a reagent, replace it.
• 70% EtOH: gallon jug for making stock is under the sink. 95% bottles are in the cold room.
• Kim wipes, gloves and Falcon tubes are located in the supply closet at the end of the hall near the rear exit.

4. General Maintenance : • Water bath: There is a monthly schedule for cleaning the water bath. Each month one lucky person gets to keep the water bath clean for the whole month! Algicide is located on shelf behind water bath.

• Microscopes: To preserve the bulbs, turn off the microscope after you use it. In order to reduce dust, be sure to cover cylinders.
• Vacuum Traps: BEFORE it is completely full, add bleach and empty into the sink. Wash out and add bleach to bottom.
• Sharps: Put coveredll containers in the hall for pickup.
• Biohazards: Replace bags when full. Stickers for pick-up are located in the top drawer. Replacement bags and ties are by the –80 freezer.
• Water pans in Incubators – you are responsible for your incubator. When low, replace with either the double distilled water by the sink or sterile water from upstairs.
• Hoods: Upon exiting, turn off the vacuum and pipettor and wipe down the hood surface with 70% Ethanol.
• Coulter Counter/Hemacytometers: CLEAN after use and turn off/put away.

5. Aliquoting reagents : (Backups of PenStrep, Glutamine and Vitamins are in the freezer in Room 225.)

• Trypsin needs to be aliquoted within a few hours of thawing. Aliquot into 2054 tubes in 1 and 2.5ml sizes. Order when you thaw the second to last bottle.
• Glutamine, Vitamins and Pen-Strep can be aliquoted within a day of thawing provided that bottle has been at 4 o after thawing. Aliquot 5ml into appropriately labeled 15 ml tubes. Order when there are 3 or less bottles left. Glutamine lasts one month in the media, so must be replenished if the media is older than that.
• FBS is normally heat inactivated and aliquoted by the person in charge of the TC room.

If you need to do this, it is very simple:

The stock bottles of FBS are kept in the basement freezer room (E17-009A). Your 101L key works for this room. The current lot Hyclone ALE13849. Thaw the FBS at 37 o in a water bath whose water level is up to the top of the serum. After thawing completely, swirl to mix the contents completely. Raise the water bath temperature to 56 o and set a timer for 40 minutes. It takes approximately 10 minutes for the water bath to get to 56 o and the heat inactivation is for 30 minutes.

Finally, if you are going to be out of the hood for > 30', please remove your supplies. Otherwise, leave a note saying who you are and when you will be back.

5. Keeping the lab clean:

Everyone should have a place to store their pipettes, reagents, and plasticware. If you do not, please see Jane. If you have items that need to be stored on the bench for any reason, you must label them with your initials. This is also true for plasticware. Any items left on the bench without identifying marks will be thrown out. We have been having difficulty with dust in the microscopes in the TC room. We will be getting the microscopes cleaned but in order to minimize the problem in the future, please make sure the cylinder has a cover if it does not have the camera attached.

6. Incubator Care:

The Forma incubators use water in a tray inside the incubator to humidify. It should be a habit to check water levels anytime you open the incubator (for any incubator). It is everyone's job who uses those incubators to keep the trays filled with water, either the double distilled water by the sink or sterile water from upstairs.

7. Hood care:

When you are finished:

• turn off the vacuum and pipettor and wipe down the hood surface with 70% Ethanol.
  
• Check the traps under the hood. If the trap is full, bring it to the sink, add bleach, wait for the bleach to work, i.e. change color and empty into the sink. Add some new bleach to the trap and put it back under the hood.
  
• Report burnt out lights, vacuum trap issues and broken pipettors.

8. TC Microscopes: Please turn off the scopes when you are finished. The bulbs last longer this way.

9. Balances:

• Please keep the balances and attendant areas clean. This means using the brush to brush off particles on the weighing plate and cleaning the area around the balance.
  
• Please turn off the balances when you are finished with them, If you are in the middle of using a balance, leave a note so that someone doesn't turn it off and you lose your tared weight.
  
• Please wash your spatulas and return them to the beaker by the balances. Disposable wooden tongue depressors are in the drawer below the balances.
  
• If you use the old balance and it's tight to turn, it needs to be cleaned. This is straightforward. Take the hanging weigh plate off the hanger. Take out the sensor and put it on the bench on its side. Take a cotton swab and dip it in 100% ETOH and swab the hole. Use a dry swab to dry any residual ETOH. Replace the sensor and the plate, and you're ready to go. If you need help, ask Jane. (rule of thumb: Jane knows everything.)

9. pH Meter Rules:

• The solution inside the probe is 3M KCL. Bottles of KCL are behind the pH Meter. The probe should be plugged with the blue plug.
• The probe should be stored in pH 7.0 standard solution.
• If you've never used a pH meter before, please have an experienced lab member show you how. Basically, you need to be aware of the fragility of the probe and not place it in your solution where it may be bumped by the stirbar.
• Dont' forget, if you use the last Pasteur pipette, please replace the canister.