Other Things You May Need to Do in the Laboratory

Autoclaving: You use an autoclave a lot in the biosciences. Things like sharp containers need to be autoclaved. Autoclave at high temp for 45-1 hour. File at Material Safety Data Sheet (MSDS). Affix label to anything you autoclave with Room #, Med Micro and data.

Trash bags should go in left autoclave (make sure plastic bag does not cover blower)

Use Right autoclave for tips, bottles. 1. Put a maximum of 2 cages with your bottles into the autoclave. 2. seal vale. 3. L=fluid inside G=gravity (dry). Use cycle 1 or 4 (20-30 minutes is enough, so come back in about an hour). 4. Hit # 4 twice. 5. Pull out and let cool. 6. Tighten caps

Autoclaving garbage bags: use 37x48'' bags. make sure says autoclave bag. double line the bag so doesnt break. get ride of the iar and tie it. put bag in metal container (1 bag at a time) and put into 1st autoclave. lock. put name on sheet. Use cycle 2 or 3 (usually 3) push 2 times. let cool 1 hrs. Put sticker on bag and thow in container.

Making RPMI

1) open one bottle of RPMI 1640 (white) (Sigma-Aldrich RPMI-1460 Medium) (1 bottle for 5L) and empty into a 3500 ml beaker. Fill the beaker at least half way with water (eventually will fill to 3500 ml mark). Drop in a stir bar and place on stir plate and turn stir knob to 7.

2) Weight out proper amount of Glucose (12.g g for 5L) and addit to the solution. Next, weight out Sodium Bicarbonate (10.0g for 5L) and add to solution. Using pipeter and 25 pipes dispense Hepes buffer into suolution (25 ml for 5L). Now you can add the rest of the water. Let solution mix well.

3) Turn on the pH meter. Place into solution. Reading should be around 7.65. Add concentrated HCL drob by drob with transfer pipet until pH is between 7.20 and 7.40. Rinse the pH receptacle with nanopure water, turn off the pH meter.

4) Filer sterilize into large beakers (fill to 500 ml on filter). You can swap filter to other bottles. Label the bottles as RPMI, your initials and the date.

Preparing HANK's Balance Salt Solution

1) Take 4 small bottles of Hanks. Tap them and put into 4 l. Fill the beaker to the 3500 ml mark with media water. Drop in a stir bar and place on stir plate with stir knob to 7.

2) turn on the pH meter and place receptacle into the solution. The reading should be about 6.6. Add 1 M NaOH drop by drop using transfer pipet until you reach pH between 7.20 and 7.40. Rinse the pH receptacle with nano pure water, and turn of the pH meter.

3) Filter sterizlize into bottles by transferring filter from bottle to bottle.