Team:Heidelberg/Notebook/miMeasure/August

(Difference between revisions)

Revision as of 08:55, 26 October 2010

seeding cells for test measurements - 96 well plate for plate-reader and FACS

test of media:

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.

@@ Line 16: / Line 16: @@
-===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)
-===26/08/2010===
-transfection - 96-well plate
-===27/08/2010===
-pilot FACS and Tecan measurements (96-well plate)
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