LABORATORY DUTIES
as of February 27, 2008

(Cammie has summarized for each piece of equipment and duties.)

Responsibilities of the person in charge of the equipment include:
1. Up keep or ensuring the upkeep of the instrument
2. That everyone is trained properly in the use.
3. The person is responsible to ensure if repair have to be done that it is done. Not necessarily by the person in charge.

THIS DOES NOT EXEMPT ANYONE FROM PROPERLY CARING FOR THE EQUIPMENT.
Though if some problem has occurred they should tell the person in charge of it, even if you are not sure there is a problem.
EVERYONE MUST CLEAN UP AFTER THEMSELVES. If they have any questions, ask the person in charge.

MICROBIOLOGY

Microscope: Zeiss Phase Contrast
Everyone: Should know how to put it in phase. Ensure it is off when one is done, covered and the slide and cover slip are clean.
Person in charge Duties: Teach people how to put the scope in phase.
- Petroff-Hausser Bacterial Counter break easily will need to be ordered periodically (have one in back up - VWR)
Person in Charge: Cammie
Dissecting Scope:

Incubators: Water Jackets - 2 LTE the 39°C and 37°C
Conventional- VWR fan driven 17°C
Percival
Everyone: Check the temperature before opening the inner door when using it.
Person in Charge Duties: Fill the Water jacketed incubators with distilled water every month.
On one LTE, one control does not work. It is noted; do not attempt to change it.
Person in Charge: Cammie
Percival:

Shaker Water Bath: 2 Amerex in 68-320 colder than room temp.)
in 68-340D sn# XQ30-1076-1
Everyone: Ensure it is properly filled with distilled water, to prevent growth in the bath and to prevent corrosion. Clean if necessary.
Person in Charge Duties: Call Amerex (925) 299-0743
Known Problems: We have a rachet to change the shaking platform.
Person in Charge:

Shaker in 30°C room: New Brunswick
Everyone: ensure you use the short screw to put in the holders the long one will rub.
Person in Charge: Cammie

Plaque Counter: Two New Brunswick C110, only one in use 68-346D
- one Monostat colony counters. 81-520-000
Everyone: Clean off any liquid that drips down and throw out old plates. Keep the area neat,
- All toothpicks go in the coffee can which goes to the kitchen.
- Capillaries thrown out in a glass waste container which is autoclaved and disposed of as glass waste. Capillaries are bought in the stock room as melt point tubes open end the largest diameter.
- All plates go into the buckets which are double bagged. Autoclaving done when full. Must be validated once a month. (See Autoclaving under safety issues.)
Person in Charge Duties: Repair -we have a new light and a new number counter. Parts are ordered from New Brunswick we can do repairs.
Replacement pens for Monostat counter Lux or VWR
Known Problems: To reorder the probe together John Fucillo can help
Person in Charge: Cammie

Experimental Water Baths:
- NAPCO (large 230, small 220)
- Thermoregulator (Hakke E-52)
- Nes Lab Cold Finer IBC 4A
2 - Nes Lab Cooling Water Bath with Flour. RTE 220
1- old cooling finger
Everyone: Fill with distilled water ONLY. If your media bubbles over in the bath or it is dirty, clean the bath. Try not to use metal rack as it promotes rust and growth, however it you do use them promptly remove the metal rack so they don't rust. Disinfect using lysol or EDTA not Chlorox it oxidizes and causes rust and leaks occur which are almost impossible to fix.
Person in Charge Duties: Known Problems
Person in Charge:

Other Water baths: Each person is in charge of their own water baths.
Person in Charge: To ensure the users know to keep the bath clean and maintain it. As noted earlier disinfect only using lysol or EDTA.

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BIOCHEMISTRY

PAGE stuff:
Now Cammie uses the BioRad Criterion Gel System, can run 8 gels
(See end of this sections for the other Gel equipment)
(See the web for running the criterion gels)
Everyone: Promptly clean equipment. Make up solution when low or tell people when low.
Person in Charge: Cammie

Power Supplies:
Pharmcia EPS-3500 in cold room hi volt and hi I
1 EC hi V hi I
1 DAN KAR Electrophoresis power supply
LKB 2197 hi V hi I
EC 500 hi V hi I
Everyone: should ensure it is off when done. Make note of bad leads
Person in Charge Duties: Take care of repairs if needed.
Known Problems:
Person in Charge:

Agarose Gel: 2 Hoefer /Pharmicia mini submarine
- 1 owl Horizontal gel
- Large IBI horizontal gel
Everyone: Pour agarose gel not too hot, it will crack the caster.
Problems: Some of the leads need adapters, they are screwed in with an Alan Wrench take them out using it.
Person in Charge:

Gel Dryers: New BioRad system
Person in Charge Duties: To ensure that people know how to use it.
Known Problem: House vacuum.
Person in Charge: Jacqueline

Heat Block Units: Many insets
Everyone: Do not put water in the well - We have killed 3 because it leaks out of the housing and corroded the electrical insides.
Every one is responsible for the one on their bench.
Person in Charge: Cammie

pH Meter: -Orion Research 310 & Ross standard electrode
Everyone: To Use - uncover the hole on the electrode ensure there is enough electrode solution in it (Ross reference electrode filling solution it is an orange color ) When done cover hole and have the bulb submerged in pH 7 solution and turn off the meter.
Known problems: It cannot be manually calibrated so if the electrode goes off the electrode needs to be replaced.
Person in Charge Duties: Periodical clean the electrode. Ensure the unused electrode is still filled and cover in buffer. Properly train people to use it.
Person in Charge:

Fraction Collectors: - ISCO on top of the incubators - Cammie
- Gilson does nto change positions well needs to be babysat - Peter
- Biorad with pump for fraction collection in cold room - Welkin
Everyone: Clean when done, it is the salt that degrades the collector.
Person in Charged Duties: Repairs
Person in Charge see above

Sucrose Gradient Maker: Gradient master
Everyone: clean up when done
Person in Charged Duties:
Person in Charge:

Dark Room: 68-340 Xomat
Everyone turn X Omat and water on 20 min before use . Turn off after use if left on and the rollers are not turning the chemical will deposit and corrode the rollers. Do not use small film. If the alarm comes on to fill up just pour the chemical in the water will fill up to the line .
In Charge Order Chemical There is a different blanket P.O. every year
Developer E. M Parker 4010 DP
Fixer - 4010 FP
Known Problems: Chemical get low and are not ordered
Person in Charge: Cammie

The U .V box , Camera and 2 hoods
Everyone - Keep clean use Saran Wrap under Gels. use double gloves . Replace the film in the camera.
Person in Charged Duties: Order film
Know Problem: the Red filter on the Camera has gone AWOL
Person in Charge:

Digital Camera: Minolta - In the computer room.
Everyone: to buy film disk when need (PC Connection).
Person in Charge:

Polaroid Slide set up: (Under Claire’s bench).
Everyone: to buy film, slide mount when we are out
Person in Charge: Cammie

Microfuges: - two Eppendorf. 5412 & 5414
- Sovall MC 12V Cammie
- Rotor to fit in the Sovall super speeds holds 32 tubes
Everyone: Clean if you spill any thing it them.
Person in Charged Duties: If in it is need of repairs
Known Problems: Have been known to become radioactive in the cold room, not lately
The Sovall has had a number of problems, door not shutting correct so it will not run..
Persons in Charge: in 68-340
in 68-322

Pipetman:
Everyone - Use properly note ones that are not working right.
Person in Charged Duties: Send to Ranian for repair.
Person in Charge: each on their own

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INSTRUMENTATION

Sorvalls: - RC 5 sn 75000956 68-320
- RC 5C +
Some rotors are found in Ingram cold room
Everyone: Clean out if any thing spills; ensure the door is closed sto the refrigeration is not continuous going on.
Person in Charge Duties: Periodical turn off to melt the ice wipe it out and clean. Service 1-800-551-2121
GSA bottle can be purchased form the stock room and the SS34 bottles form VWR and Fisher.
Known problems: A Long history of problems with the RC 5
Person in Charge: Cammie

Beckman: L8-55M
Everyone: Turn on set temperature and start vacuum. It will not cool down if the vacuum, is not activated. Set speed and time, activate vacuum, the vacuum is down to 150 microns, start the run however one must push Enter/Display in order for it to begin the run. Wait till it achieves speed so you can determine if there are any problems.
Note: If you are using the SW27.1 you must only accelerate to 170 rpm as noted on the cfg Or it will become unbalance and the cfg will stop.
When finished using the cfg activate the "Dry Cycle". When done turn off. Order tubes when low. Beckman ultra clear you can get them in the stock room.
Service new company
Person in Charge:

Computer: in 68-330AA
3 G4 2 in computer room, 1 in lab on scanner
1 G4 Cindy
G3 Jon
2 titaniums Peter/lab 7 Jon
PCs on the - 2 Cary 50 Bio, Hitachi Fluorescence Spect -F4500, FPLC
- Power Book Jon, Peter and Cammie
Everyone: Keep area clean, Clean up after your self.
Person in Charge Duties: Trouble shoot
Person in Charge: MAC- Cindy
PC -

Spectrophotometer: - 2 Cary 50 Bio in 68-316 Melissa
68-346D Cammie
Everyone - Keep area clean. Remember if one uses proteases, clean the cuvette out with the EtOH and HCl solution.
Person in Charged Duties:
Known Problems:
Person in Charge: see above

Hitachi Fluorescence Spect -F4500
Everyone - Keep Area Clean People need to be trained by King people need to sign up .
We need to ask anyone we see on it who trained them and if they signed up
Known problems: Occasionally sample door does not close properly
Note: Be sure to keep track of lamp use time in the log book.
Wash cuvettes thoroughly with ddH₂O and EtOH after use and do not put them away until they have dried completely.
Do not store data on the Gateway that runs the spec, as it crashes frequently
Refractometer (on bench beside fluorimeter)
-wash quartz with ddH₂O only, wipe dry gently with Kimwipe
-after use, fold-up a Kimwipe and place between quartz to prevent scratching
Person in Charge Service - Hitachi
Person in charge:

Microplate Reader: Dynex
Everyone to keep clean
Known problems need to read how to save the data to text before running the assay.
Person in Charge: Cammie

Refrigeration: Puffer Hubberd Unitherm- 68-322 personal shelves Claire
Amana 68-346D Other Stuff
Sears Cold Spot 68-340D
Sears Cold Spot 68-320 Master stocks
Little Brown 68-346 SDS Buffers
- Raidoacitve storage refrg. in 68-346D
Everyone - Keep them neat and Clean
Person in Charge Duties: Service Shon's Refrigeration 326-1947, Delegate space in the refrig.
Person in Charge:

Individual refrigerators at benches: This is the responsibility of the person at the bench. Repair if needed is Shon's but it may be cheaper to buy at new one.

Freezer : Harris -85oC sn # S24C-140075TCK M# HLT !7LS85 -DBA inherited from Baltimore
Revco 68-322
Revco 68-340
Everyone - Store long term samples. In Cardboard boxes in the metal racks. Short term in styrofoam boxes. Don't write on boxes use tape to label ensure the styrofoam boxes are secured closed with tape or rubber bands they tend to open and you lose samples. People should clean out the samples. All radioactive samples should have radioactive tape labels.
Person in Charge Duties: Repair Shon's Refrigeration 326-1947, Order need boxes and racks.
Person in Charge:

Balance: Mettler P1200
Mettler AT 250 in 68-340E
Everyone - Clean any mess left on it or around. TURN IT OFF WHEN DONE.
Person in Charge Duties: Repair Service by Mettler 1-800-METTLER
Person in Charge Cammie

Hood: In 68-346D and 322
Everyone - Keep clean, should be used for experiments not store chemicals. Do not obstruct the back baffle, it blocks air flow.
NOTE: There is a label on the side that indicate the position of the sash to get proper air flow.
Person in Charged Duties: Unfortunately you may to clean it up. Repair
Known Problems: Becomes the dump for everything.
Person in Charge: Cammie

FPLC: Pharmaica
Everyone:
-Keep area clean
-Intake values should be stored in ddH2O
-Columns should be equilibrated in an EtOH gradient
-New users must have an account created by a system administrator
(Melissa or Steve R.)
Known problems: Current software manual is for Unicorn 3.2, but the software is Unicorn 4.0
Contact: Tim Rice (Amersham Pharmacia)
Person in Charge:

Thermocycler: MJ Mini Cycler
Everyone
Person in Charge:

Electron Microscope
Person in Charge:

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LAB ADMINISTRATION

Master Stocks: In the White Refig in 68-320
Everyone - Are to be used for an occasional experiment. If one is going to use it more than once, the master stocks should be used to isolate the plaque to grow your stocks, so there will be one original.
If the stock is low, tell Cammie so she can ensure a new stock is grown. The stocks should be tested to guarantee it still contain its allele.
Known Problems: Most of the stocks are low titer and may not be have enough to do an experiment, therefore you may need to make one up.
Add new stock to the Computer Stock list on the Mac in File Maker Pro.
Person in Charge: Cammie

Bacterial Strains: In the Freezer
Everyone - To tell person if the stains are no longer active. Add strains not in the collection.
Peter has DMSO in sterile vials mix older overnight cells and freeze at –20ºC
Person in Charged Duties: To ensure the strains are functional and to put in new strains. Instruct people how to add to the collection
Person in Charge: / Cammie

Internal Ordering
Everyone: When going to the stock room pick up all that is on the list in 68-330aa. Can use the credit card.
Outside ordering: Cammie has a File Maker Pro to keep your orders on the computer room computers. Make a print out if credit card & put it in the credit card folder. When the packages come in, check the packing slip against the contains of the box, sign it with a date. Place in Cindy's black metal slot or Visa folder if credit card purchase. (Attach the delivery slip; this is useful if something has to be sent back under warrantee.)

Chemical Shelves: Stored in alphabetical order except for common reagents which are stored above the balance.
Everyone - If we are low on a chemical put it on the shopping list.
Person in Charged Duties: to periodically make sure the chemicals are still in order.
Person in Charge: Everyone

Recycling: check the MIT recycling page for any questions.
The grey waste baskets. Paper only in the office, computer room:
White, colored, glossy & magazines, newspapers, catalogues, envelopes w/ plastic windows, manila envelopes (remove metal clasp), file folders, small pieces of cardboard.
No food wrappers or trash in recycling bins!!

Everyone - Promptly clean equipment. Make up solution when low or tell people when low.
Person in Charged Duties: to order supplies which includes glass plates, spacer, gel boxes, staining boxes. Order new plates and spacers when needed,
- 15 cm x 16 cm larger plates from Dan Kart or Owl,
- 0.75 mm spacer and combs from Hoeffer.
- Mini gels are from Owl thus the replacement supply might come from Owl
- Bio Rad Mini Gel can be gradient or 71/2%
Known Problems: Some of the old gel boxes leak can be fixed with methylene chloride or chloroform ,or propylene oxide.
Person in Charge as of Sept.98:

Group Meeting: To ensure people know a week ahead they are talking and to remind people they are bringing food. To determine if it is feasible to have group meeting due to lack of attendance because people are away.
Person in Charge - Cammie

We have but not in use:
Sequencing Apparatus: 2 Gibco BRL apparatus
Everyone - Keep clean do not use grease it is messy
Person in Charge as of Sept.98:

Condenser & Chemical Trap: Savant Refrigerated Condensation Trap model RT-100A
Everyone - Turn it off at the end of the day and empty the next morning or the liquid gets to the level that it blocks the inlet hole and will not pull a vacuum. Ensure there is EtOH in the cooling cavity.

NOTE: The tubing has hook up instructions on them. The vacuum pull from the port towards the top and the "dryer liquid" goes to the bottom. Or you will be pulling the acetic acid into the neutralizing trap shorten its life. The trap is placed between the pump and the Condenser.

Person in Charged Duties: Order new trap when the it breaks and Neutralization cartridges, when it has become saturated.
- Model D.C 120 A Disposable Cartridge Acid Vapor Neutralization $49.00
- Glass trap GIT-100 - about $120.00.
Savant Instruments 1-800-634-8886
Known Problems: As described above, freezing up of the trap because not emptied. The improper hook up of the hoses.
Person in Charge as of Sept.98:

Speed Vac Concentrator: Savant
Everyone - Ensure it remains clean.
Person in Charged Duties: Periodical grease the O-ring.
Person in Charge as of 98:

LKB Isoelectrofocusing Unit
Not used that often lent out
Person in Charged Duties: to know who has borrowed it.
Known Problems: "dead in the water"
Person in Charge as of 98: Cammie

SDS Gel stuff: 2 Owl double 15 cm x 16 cm, .75 mm spacers with notched plates
- 2 Dan Kar double boxes for the large 15 cm x 16 cm gels
- 2 Owl single Boxes
- 2 Mini Owl 4in x 4.5 in 1.5 mm spacer.
- 2 mini penguin
- 2 BioRad Mini
- 1 transfer for the BioRad Mini
-1 Hoeffer/Pharmcia Transfer apparatus Larger TE 42

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