The assay was first performed in L-15 containing 0.6% low melting point agarose and 10% FBS. We seeded 1000 cells in 1 ml of L-15 containing 10% FBS and 0.3% low melting point agarose (Amresco, Cleveland, OH, USA) and laminated it to the substrate. Two weeks later, the plates were stained with 0.2% crystal violet and the number of cell colonies was counted under an optical microscope (IX70; Olympus Corporation, Tokyo, Japan) [19 (link),20 (link)].
Protocol detail
Soft Agar Colony Formation Assay
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Agarose
Assay
Cells l
Crystal violet
Optical microscope
Corresponding Organization :
Other organizations : People's Hospital of Shiyan, Hubei University of Medicine
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Variable analysis
independent variables
- Concentration of low melting point agarose (0.3% vs. 0.6%)
- Number of cell colonies
- Cell line (not specified)
- Seeding density (1000 cells/ml)
- Culture medium (L-15)
- Fetal bovine serum (FBS) concentration (10%)
- Staining method (0.2% crystal violet)
- Microscope used (IX70; Olympus Corporation)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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