finkbeiner lab

Descriptions of jobs for the Finkbeiner Lab:

†

Click on the Link Below to see Full Descriptions:

1. Delivery Receving Person
2. Working Solutions for Neuronal Cell Culture
3. Neuronal Cell Culture Stocks
4. General Cell Culture Stocks
5. Bacterial Culture Stocks
6. Tissue Culture Upkeep and Supplies
7. General Lab Supplies and Equipment Maintainance/Repair

Delivery Receving Person

1) Unpack and/or deliver items that have arrived to lab members or place them in newly arrived containers (boxes in the cold room (4C), first -20C freezer on the left in the hall, and in the bottom shelf of the -80C). This includes placing media in the cold room and tissue culture supplies in the appropriate storage spaces.

2) Check off items that have arrived in the ordering binder. The binder is next to the centrifuge in the first bay on the left as you enter Room 243.

Generally, deliveries are dropped off twice a day in the first bay on the left as you enter room 243.

If you are unavailable at the time that deliveries come in, make sure that orders binder is updated (check with other lab members) - items that other people have unpacked and stored should be initialled as arrived.

Working Solutions for Neuronal Cell Culture

Check with lab members to determine the usual or unusual volume needed per week of these solutions, making sure that there is always enough:

1) Dissociation Medium

2) 10x KY

3) 2x Sterile HBS

The recipe for making each of these is available in the Neuronal Culture protocol.

Check that we have at least 10-12 bottles of BME, DMEM, and Optimem in the cold room. If not, write up the necessary amount on the white board. If we are low on aliquots of stock solutions, inform the person responsible for them.

Lab Stocks - Neuronal Cell Culture

1) SVM, 1.5, ITS, Serum, Putrescine, Transferrin, progesterone, Hyclone Bovine Calf Serum (current lot# ALE13644)-See Primary Culture Protocol for Complete list.

2) If we are low on pH"ed 2x HBS for primary culture transfections, talk to Montse/John about how to make more

2) If more of any component needs to be ordered, put request on white board.

Lab Stocks - General Cell Culture

1)†††† Aliquot:

a.†††††† 100x L-glutamine (5-10ml aliquots)

b.††††† 100x pen-strep (5-10 ml aliquots)

c.†††††† 4% PFA (12ml aliquots)

d.††††† 100x Glutamaxx (5-10ml aliquots)

e.†††††0.25% Trypsin Solution (5-10ml aliquots)

f.†††††††Gibco Calf Serum (50ml aliquots)

g.†††††Gibco Fetal Bovine Serum (9 ml aliquots)

h.†††††Hyclone Equine Serum (HC ES; good lots: AGL7527; ALK14933, AJG10637) (50 ml aliquots)

i.††††††††Hyclone Supplemented Calf Serum (HC CS+; good lots: AKH12338, AMC15812) (50 ml aliquots)

2) If fewer than 6-8 aliquots remain, put request on white board and include lot number.

Lab Stocks - Bacterial Culture

Maintain Stocks of Bacterial Antibiotics: Amp, Kan, (Cam)

Maintain a plentiful number of LB-agar plates with:

1)††††Amp

2)††††Kan

3)††††No antibiotic

If no LB is present, make some up and autoclave it.† Consolidate LB into as few bottles as possible.

If reagents for making stocks are in short supply, add item to white board to be ordered.

Tissue Culture Upkeep and Supplies

Fill Finkbeiner 70% Ethanol spray bottles

Make sure that overfilled biohazard bags are put in the containers out in the hall and replace them with empty bags if some inconsiderate lab member has forgotten to do so. If the vacuum traps are full, add some bleach and empty.

Clean hoods thoroughly at least once a week: Wipe down all surfaces with dilute bleach, then with EtOH. Empty waste traps and rinse them with bleach.

The following is largely taken care of John Gray, but if anything needs to be done, please do so:

Make sure shelves next to both Finkbeiner Tissue Culture Hoods are stocked with plates/syringes/pipettes

Make sure tissue culture supplies are fully stocked in neurophysiology: 35, 60, 100, 150mm tissue culture plates, 6, 12, 24, 96 well TC plates, 2, 5, 10, 25 ml disposable serological pipettes, 1, 5, 10, 30 ml Syringes, luer lock 0.2 um filters, 0.2 um bottle top filters (45 mm), 15, 50 ml conical tubes, 5, 14ml round-bottom tubes, T75 flasks, etc. There should always be one unopened box of everything present - all labeled shelf space for supplies should be filled. If not enough of any item is present, talk to John Gray and, if necessary, place items on white board for ordering.

Equipment Maintenance/Repair

Contact Service Representatives for Equipment that is mal (non) functioning. Place Notes on Equipment when it is not functioning properly, including when the equipment is likely to be usable again.