RGG Cleanlab routines

Norwegian College for Fishery Science Research Group for Genetics Last updated: November 2019 CLEAN LAB ROUTINES – C358 BEFORE ENTERING THE CLEAN LABS You must be freshly showered and wear a clean set of clothes, have entered the NFH-building so that you avoid certain high-risk contamination areas. You must enter the NFH building from the main entrance or the east side entrance (by the new building). When entering the lab area, you must only enter in the lab corridors from the door between stairs and elevator (opposite from lab B310). The high-risk contamination areas are our genetic lab B310 and adjacent rooms and corridors, and ‘Fiskemottak’ area on the first floor and adjacent rooms and corridors. Finally, you cannot have eaten any fish on the day of entry. Firstly, before you enter the lab, do you have everything you need to perform the lab task at hand? We do not want you to leave in the middle of the process. Remember to visit the restroom as well. GENERAL CLEAN LAB ROUTINES 1. Take your shoes off in the corridor and bring with you the plastic box (if there is any stuff in it) to bring into the labs. Enter the lock. 2. Leave personal belonging in the small box (i.e. phone, access card, rings etc.) Wash your hands, put on clean suit and blue shoe covers, and finally a pair of gloves. 3. Make fresh 10% bleach in the bucket, and more 50% EtOH solution in the spray bottles if needed. Take a new dish cloth. 4. Clean what was in the plastic box from the corridor and place them where it belongs. 5. If you have samples to bring into a clean lab: a. Remove the outer bag and clean the next bag with bleach before entering room. If your samples are not double-bagged (which means only one bag around you triplicates samples), then you clean the bag very thoroughly with bleach, but not ethanol since it will wipe off the label. b. Enter the room with only the bags of samples. Leave the sample box in the lock. Put samples in another box inside the extraction room box and put in the fridge. c. Exit back to the lock. Clean box used for samples and also the box from the corridor. d. Change gloves. Re-enter the extraction room with bleach, ethanol and lab equipment you may wish to bring in. 6. When inside the extraction room put on lab shoes. Put your lab equipment in assigned box with project name. Keep all your things in this box always and keep it clean. 7. Clean you gloves and put a second pair on. 8. Clean the flow hood and all equipment that are to be used for the protocol, i.e. vortex, centrifuges, pipettes, racks on so forth. Clean the orange 50ml tube adapters from the centrifuge and place inside the flowhood. Clean the centrifuge cups. Always clean everything. 9. Place everything in the flow hood and UV treat it for 10 mins. At this step you can even place the tubes, tips, bags, markers etc. needed for the lab protocol in the hood to be UV’ed too. NEVER UV the chemicals from the kits. 2 10. While waiting for the UV to finish, first clean the heating cabinet and switch it on, the proceed to clean the large falcon centrifuge, sample wheel (rotator), chair and any other surfaces. Put bags in the two trash cans – one for regular waste and one for lab waste. 11. Change the second pair of gloves and clean them. 12. Proceed to the appropriate lab protocol. When extractions are done, your samples should be stored in the stock and aliquot freezers respectively. Remember, do not freeze your aliquots if they are to be used within 4 weeks for downstream protocol (i.e. lib.prep., PCR etc.). Store in fridge if that is the case. Store your aliquots in PCR plates or strips. Put two bags around and make sure to label it properly. The rest of your extracted DNA (stock) is stored in 1.5 ml tubes in a cryobox that you have purchased at the store and brought with you. Make sure to label the box properly. Put the cryobox in two bags before storing it in the stock freezer. AFTER LAB TIME AND EXITING CLEAN LAB 1. Clean your own stuff and put back in assigned box. Clean the sample box that you used for your samples in in the fridge. 2. Clean the flow hood and all equipment that has been used, i.e. racks, vortex, centrifuges, pipettes. Clean everything! a. Use the same rack always and keep it in your box of lab stuff. When the project is done, then you bring it out to the slues and give it a proper wash/cleaning. 3. Place everything back in the flow hood and UV treat it for 30 mins. 4. Clean the large falcon centrifuge, chair and all other table surfaces. 5. Empty both trash bins and put in new bags. You will take both bags with you out of the room when you are completely done. 6. Go through the checklist before you leave. 7. Take off lab shoes and exit to the lock. Bring with you the bleach bucket, ethanol bottle, MilliQ bottle and the trash. 8. Bag with lab trash goes in the yellow bin, the other you take with you out of the lab. 9. Empty bucket with bleach. Throw out the dish cloth in regular trash. Put bucket, MilliQ and ethanol spray bottles back where they belong. 10. Take off the blue shoe covers and throw them out in regular trash. Take off your clean suit and put in bag with you name. Place in bookcase. 11. If needed, empty the regular trash bin in the lock and put in new bag. If a yellow bin is full, put lid on. Bring trash, yellow bin and box from the corridor out with you. 12. Leave the trash in the hallway for the short time while you go to the store to get the things needed for your lab protocol or the lab. Remember to not pass any high-risk areas. In case the store is closed when you are finished you can buy the necessary things the next morning. 13. Label the things that are yours and put all things in the box in the corridor to bring in one of the next days. 14. If you are not to re-enter the clean lab the rest of the day bring the trash to lab B310. Remember to fill out blue label for the yellow bin. Ask Julie if you have any questions. Remember to shower and fresh clean clothes on the day you wish to work in the clean lab. It is possible to shower at the NFH-building right next to the SIMFISH-meeting room on floor 0.