Developmental stage: early stationary phase Tissue: cell suspension
Treatment protocol
20 ml of an early stationary phase MM2d cell suspension (7 days after previous subculture) was transferred into 100 ml fresh MSS-medium (MS-salt, 3% sucrose, 0.5mg/l NAA, 0.05mg/l kinetin) and incubated in the presence of 4.16 ug/ml aphidicolin (not exp.variable) for 24 hours under continuous darkness by rotating at 130 rpm at a temperature of 27C. Cells were gently washed with 1 l MSS-medium through a nylon net (47um mesh size), followed by centrifugation (387 x g for 1 min, no brake applied) to remove the drug. Cells were resuspended in a total volume of 120 ml fresh MSS-medium, incubated as above and samples were taken 2-3 hourly for analysis. The T16 sample was taken at 16 hours after washing to release the block (exp. variable; time course).
Extracted molecule
total RNA
Label
biotin
Description
DNA replication inhibitor: aphidicolin growth media type: liquid culture media light regime: continuous darkness nutrients: 100 ml MS salts, 3% sucrose, 0.5 mg/l NAA, 0.05 mg/l kinetin average daily temperature: 27oC
Data processing
The data were analysed using Affymetrix Microarray Suite 5.0 (MAS5)
