Preparing rotation medium
From mn.fysikk.laglivlab
Contents
Stock protocols
Stock sugar water
(Done at the Giaever lab)
- Measure up 3,33 grams sugar (farin/sucrose)
- Fill a 15 ml tube with 10mL distilled water
- Add sugar to distilled water and shake
EDTA working solution protocol
- Fill a 15 ml tube with 2,25 ml distilled water
- Add 250 microliters EDTA from stock solution to the distilled water in the 15 ml tube
BSA working solution protocol
(This preparation should be done in the clean lab biosafety cabinet)
- Fill a 15mL bottle with 5mL distilled water
- Slowly add 0,5g BSA powder to the 5mL distilled water whilst slowly stirring
- Store at 4 celcius
Protocol dilution sequence
(The first 5 steps require the use of a bio safety cabinet and must be done in the clean lab. all steps after step 5 can be done in the Giaever lab)
- Remove 1mL GM with diluted cells
- Centrifuge the GM at 400 RpM for 4 minutes
- Remove the supernatant without disturbing the pellet
- Add 29 microliters RPMI to the 15ml tube containing the cell pellet
- Add 40 microliters of BSA working solution to the 15mL tube containing the pellet
- Add 1mL sugar water mix (33% w/v) (dilute 0,33 g sugar in 1 mL water)
- Quickly add 40 microliters EDTA working solution
- Quickly fill the tube with distilled water until the solution has a total volume of 4 ml and shake gently