1. Klebba Lab Rules and Regulations July 2004

2. Golden Rules If you open it, CLOSE IT If you turn it on, TURN IT OFF If you break it, REPORT THE BREAKAGE TO MARJ, PHIL OR SALLY If you borrow it, RETURN IT If you make a mess, CLEAN IT UP If you move it, PUT IT BACK If you use it all, DO NOT REPLACE empty containers: INFORM MARJ WEAR SAFETY GLASSES WHENEVER NECESSARY REDUCE GLOVE USAGE WHEN POSSIBLE

3. Cleaning & disposal Sink area: only materials to be washed; not contaminated by bacteria Plates: in orange bags that will be autoclaved before disposal Contaminated glassware: in plastic buckets NO GLASS IN NORMAL TRASH CANS TOXIC CHEMICAL WASTE: dispose in appropriate containers & LABEL

4. Media, buffers, solutions Plan your experiments in advance; check how many plates will be needed. Inform Marj if only a few plates are left in the cold room If you need to use more than 500ml liquid media, inform Marj Many buffers are ready in 10X form for common use (TAE, PBS, blocking buffer for western) Special solutions and buffers: MAKE YOUR OWN.

5. SDS-PAGE Wear gloves when handling plates, acrylamide is VERY toxic – rinse pipettes with water before placing in the wash; remove polyacrylamide from flasks Avoid storing more than 2 gels at a time Spacers and combs should be placed back in the drawer after use – do not leave them by the sink! Make Lower Buffer, Upper Buffer & Acrylamide/Bis if necessary – composition is written on the bottle When preparing acrylamide/bis, wear gloves and a mask, wash the balance area when finished Rinse apparatus with water right after use – salts are corrosive Turn power supplies off and store electric cords away

6. Western blotting Minimize use of membrane For one gel: 100mA - 2 hours RINSE APPARATUS, and store it in the cabinet Antibodies can be used a few times: do not discard cocktail after a single use; add azide, LABEL and keep in the refrigerator for up to 5 experiments.

7. AGAROSE GELS Storage is OK, but use common sense: make only what you will use in the near future DO NOT LEAVE AGAROSE inside flasks, rinse out while warm Change the buffer in the gel box every week and write it on top of the lid – WASH THE BOX BEFORE CHANGING THE BUFFER DNA Ladder is expensive: do not use it in large wells, apply it to a small well in a separate gel running next to it

8. BALANCE Clean every spill you make, no matter how small Put chemicals back unless the bottle is empty Be very careful with toxic chemicals

9. Electroporation Cuvettes are expensive Rinse cuvettes with water before putting them in the bottom drawer Remove ALL your stuff from the electroporation area: tips, eppendorfs, ice Turn OFF electroporator: switch on and off 4-5 times

10. SPECTOPHOTOMETER Log your use of the UV lamp: TURN IT OFF after use Quartz cuvettes are VERY expensive and VERY delicate: handle with care, wash and store – do not leave them upside down in the washer, they might fall and break EMPTY THE WASHER

11. COMMON AREAS Clean electrophoresis spaces after you use it (agarose & protein) Label any leftover buffer Clean UV light box after each use Clean magnetic stirrer/heater: TURN IT OFF when you are done using it Remove markings from centrifuge tubes

12. Your work area CLEAN YOUR BENCH EVERY DAY! Discard your ice at the end of the day Discard old, contaminated plates and cultures Do not leave samples in ice overnight, so that next morning it’s all floating in water. Put your samples away and store the ice. Your work area reflects the kind of professional you are: make it shine