seeding cells for test measurements - 96 well plate for plate-reader and FACS
test of media:
OptiMEM 0% FBS + Pen/Strep + L-Glu + other according to cell line
OptiMEM 2% FBS + Pen/Strep + L-Glu + other according to cell line
DMEM 10% FBS+ Pen/Strep + L-Glu + other according to cell line
cells did not adhere properly so we decided to coat 96-well plate with poly-L-lysine and repeat
significantly better growth in DMEM with 10% FBS, so we decided to grow cells in it and before measurement to change it to OptiMEM, to reduce high background fluorescence coming from FBS.
09/08/2010
seeding and transfecting cells for microscopy test measurements
add 0.6ul FuGENE reagent to 20ul of OptiMEM
mix and incubate 5' at RT
add 0.2ug DNA
mix and incubate 15' at RT
add DNA-FuGENE solution to 10 000 cells (400ul)
mix and incubate 10' 37degC 300rpm (shaker for 15ml falcon tubes is in the cell culture room at 3rd floor)
transfere cells to the plate
grow 48h
10/08/2010
coating plates with poly-L-lysine
add 15ul of poly-L-lysine solution to each well (make sure whole surface is covered with solution)
leave for 30' in the incubator
remove poly-L-lysine solution
wash once with PBS
new 96-well plate was prepared (wells 2500 cells Huh7 are contaminated with HeLa) like previously
25/08/2010
seeding cells for pilot FACS and Tecan measurements (96-well plate)
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