Instructions for working in Majids category 2 hood

Plan your work and liase with Amelia () to check that the hood will be free for your use.

Liase with designated person to collect samples

Set a time which would have allowed you to set up the hood in advance but still allow time for you to do your work and tidy up (suggest you try about 10-11am for first go)

Gather together beforehand

The tips and tubes you will need, wash bottle for virkon. A spray bottle for IMS, autoclave bag and box, gloves, seal-able container(s) for transporting samples and seal-able container for waste within the hood, pipettes, K dry, lids etc and calculate the amount of solvent needed. Also pack a spill kit in case you should need it. (see separate sheet for spillages)

The morning of your work

Make up a 2% solution of Virkon (suggest 1 litre volume) mix well, half fill a wash bottle, half fill a sealable pot (for waste later)

Fill the spray bottle with 70%IMS.

Take out the solvent required for your experiment.

Place everything you need in a carrying tray with handle (so you can open doors easily) and go upstairs. You may need two trips to bring up autoclave bag and box.

When ready to set up in lab CRPC5.23

Turn on light switch to room from outside door.

Turn on vent on wall (there is CO2 cylinders in the room)

Put a clean lab coat on

Turn on the catagory 2 hood, two switches at top whilst the front plate is still in place.

Remove plate and place by door out of the way.

Wearing gloves, spray hands with 70% IMS and proceed to spray around work area and wipe out hood first with IMS then Virkon. Place soiled paper towels in our autoclave bag.

From now on anything placed in the hood must first be sprayed with 70% IMS including your gloved hands this is to protect Majids future work with cell cultures.

Place in hood our pot with 2% Virkon, pipettes tips and tubes.

Extract our tissue as per method. (see separate sheet)

Remember if working for a long time in the room there is air conditioning, use it. Door to room is to be kept closed whilst you are working.

At end of work place all your solvent tubes in seal-able pot for transport downstairs.

Wipe down pipettes with IMS before removing from hood, pack your stuff away into transporting tray. Wipe out hood with virkon and IMS placing all

soiled tissues and gloves in autoclave bag. For moment leave bag and pipette tip pot upstairs. ( tip pot - seal with lid and shake gently so that all tips are under the surface of the virkon, then leave for 24hrs to soak)

Once hood is clear of everything except waste pot replace front plate BEFORE turning off hood.

Turn off hood (two switches on top). Turn of vent (and air conditioning if used).

Turn off light as you leave room.

Once downstairs in ‘our’ lab double check your tubes, place foil under caps check labelling and store in freezer.

Tell the designated person what you have done so they can alter the database accordingly.

Any remaining tissue not exposed to solvent, That is if you have split a sample into pieces MUST BE RETURNED IMMEDIATELY TO THE DESIGNATED HUMAN TISSUE FREEZER.

REMINDER IT IS ONLY SOLVENT EXTRACTED TISSUE THAT CAN BE HELD IN CRPC LAB 404.

Next day

Go up to Majids lab taking spray bottle of 70% IMS and paper towels and turn culture hood on, wearing gloves grab the tip pot, once again spay and clean out hood which should now be completely empty of items. Turn hood off. Take tip pot and autoclave bag down to ‘our lab’

Do not open doors or walk around building with gloves on. Use a fresh pair in the two different labs.

Once everything is back down in Liz’s lab strain tips from liquid layer and put tips in autoclave bags. (Liquid goes down fume hood sink with plenty of water)

Take autoclave bag in box up to Lynn for disposal. Place in her metal container for autoclaving. Bring cardboard box back for later use. Do this immediately on return to lab.

Draft July 09