Klebba Lab Rules and Regulations

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