Maloof Lab:Undergrad Instructions

1. Put clean dishes away: ~5 minutes depending on previous load of dishes.

     (Once or twice a week depending on volume of dishes needed to be washed.) 

1. Open dishwasher.

2. Put glassware and washed items back into glassware shelf.

2. Clean dishes: ~5 minutes depending on amount of dishes needed for washing

     (Once or twice a week depending on volume of dishes needed to be washed.) 

1. Check amount of dishes needed to be washed in plastic tray.

2. Handwash graduated cylinders and tube racks with dish soap.

3. If they have marker marks on them, use ethanol to wash off.

4. Peel off all tape on glassware and caps before putting into dishwasher.

5. Place caps, spatulas, stir bars into cage.

6. Place other items where appropriate.

7. Place pestles in empty glassware.

8. Put detergent in and turn to power scrubber setting.

3. Check trash-autoclave:

     ~5 minutes to gather trash and load into autoclave/replace bag
                 Autoclave: 30 minutes
                 Total time: 35 minutes
     (Once or twice a week depending on volume of trash.) 

1. Check level of trash

2. If getting full, spin the autoclave bag opening into a knot and use autoclave tape to tie around the opening. Then slap on a small piece of autoclave tape on bag as well.

3. Put into plastic autoclave tray.

4. NOTE: autoclave fits a large plastic tray and small plastic tray simultaneously. NEVER PUT AUTOCLAVE TRASH AND STERILING ITEMS IN SAME TRAY!

5. Sign up for autoclave machine and put it in. Set at 121 degrees Celsius, Wrapped cycle at 30 minutes. Make sure the sides of the autoclave bag do not touch the side of the autoclave. Press start.

6. Replace old autoclave trash bag with new bag in the now unbagged autoclave trash bin.

7. Come back after 35 minutes and collect trash with autoclave gloves on. Throw away autoclaved trash into trash bin. Bring back the autoclave tray.

4. Make tips: Put them in box: ~10 minutes to put in box and stick autoclave tape on

                              Autoclave: 30 minutes
                                   Dry: 25 minutes
                             Restore in cabinet: 5 minutes
                 Total Time: 1 hour 10 minutes
     (Once every 2-3 weeks) 

1. Check level of tips available. If low, proceed to second step.

2. Gather appropriate tips in stock room. Rip plastic off.

3. Gather empty tip boxes and put new tips into tip boxes.

4. Put autoclave tape on each tip box. Then put all tip boxes into plastic autoclave tray.

5. Sign up for autoclave and set wash cycle at 30 minutes. Exhaust at 25 minutes. Wrapped Cycle. 121 degrees Celsius.

6. Come back after an hour and put the newly autoclaved tips into tip cabinet.

5. Autoclave tubes: Put them in jar: 5 minutes

                              Autoclave: 30 minutes
                                   Dry: 25 minutes
                 Total Time: 1 hour
     (Once every 5-10 weeks)

1. Check tubes level. If running lower than two jars each (four total), then proceed to step 2.

2. Gather tubes into plastic jar, piling on different colors over each layer.

3. Put aluminum foil over the jar. Place autoclave tape on top.

4. Sign up and put into autoclave. Set at Wrapped cycle, 30 minutes wash, 25 minutes exhaust, and 121 degrees Celsius. Put into plastic autoclave tray.

5. Come back after one hour and put the newly autoclaved tubes onto shelf.

6. Check Miliq Water: 5 minutes upward to 15 minutes depending on amount of water left

     (Daily) 

1. Check Miliq water level. Carry jug into other room.

2. Press the recirculating button until reading reads 18-18.2.

3. Turn to dispense with dial after putting the tube into the jug. Wait until fully filled.

4. Turn off the dispense dial and turn off the reciruclating button. Bring jug back.

5. Untwist gap slightly to allow easier dispensing for others.

7. Fill TAE and SDS: 5-10 minutes depending on amount of TAE or SDS left.

     (Daily) 

1. Check level of TAE and SDS. If running low, proceed to second step.

2. If running low, put in 50 mL of 50x TAE or SDS with 2450 of distilled water.

3. Untwist cap gently to allow easier flow when dispensing for others.

  To make 50x TAE:
   Into 750mL MilliQ water add:
   242 g Tris base
   57.1 mL Glacial Acetic Acid
   100 mL 0.5M EDTA
   Adjust the final volume to 1L (1000 mL) with MilliQ water.  

8. Make LB, LB Agar, 2X LB, 2X Agar:

     15-20 minutes to prepare in glassware
     Autoclave: 25 minutes of active use + 10 minutes of cool down
     Total Time: ~1 hour
     (Once every 5-8 weeks depending on supply) 

1. Measure appropriate amount written on bottle for item that is running low on balance.

2. Put in appropriate amount of Miliq water in glassware. Then put the particular measured amount into glassware. Stir well unless it is agar.

3. Clean balance area.

4. Put cap onto glassware. DO NOT TWIST ALL THE WAY CLOSED!! UNTWIST TWO TURNS TO ALLOW CAP TO WOBBLE AND NOT BE COMPLETELY CLOSED!!

5. Place autoclave tape onto cap.

6. Label item name, your initials, and the date you made it onto an ordinary tape and place on the glassware for identification.

7. Place into plastic autoclave tray.

8. Sign up for autoclave and place into autoclave on liquid cycle, 25 minutes, 121 degrees Celsius.

9. Come back in 35 minutes and put newly autoclaved liquids into shelf. If agar, mix well before placing in shelf. TWIST CAP FULLY CLOSED NOW.

9. Autoclave Miliq water:

                 Prepare in glassware: 5 minutes
                 Autoclave 25 minutes of active use + 10 minutes of cool down
                 Total Time: ~45 minutes
     (Once every 3-4 weeks depending on supply) 

1. Measure 200 mL of Miliq water into glassware.

2. Put cap onto glassware. DO NOT TWIST ALL THE WAY CLOSED!! UNTWIST TWO TURNS TO ALLOW CAP TO WOBBLE AND NOT BE COMPLETELY CLOSED!!

3. Put cap onto glassware. DO NOT TWIST ALL THE WAY CLOSED!! UNTWIST TWO TURNS TO ALLOW CAP TO WOBBLE AND NOT BE COMPLETELY CLOSED!!

4. Place autoclave tape onto cap.

5. Label item name as “200 mL Sterile Miliq H20”, your initials, and the date you made it onto an ordinary tape and place on the glassware for identification.

6. Place into plastic autoclave tray.

7. Sign up for autoclave and place into autoclave on liquid cycle, 25 minutes, 121 degrees Celsius.

8. Come back in 35 minutes and put newly autoclaved liquids into shelf.

10. Other activities upon needed: variable