Two hundred to 500 cells/well were plated in 6-well plates overnight. Cells were treated with either single or combination treatment for 5 days after which the media was replaced with regular media without drugs. Colonies were allowed to form for an additional 7 days, with the media being replaced every 3 days. Cells were fixed after 12 days for 20 min using 4% buffered formaldehyde. Colonies were stained with 0.05% crystal violet solution for 10 min following two washes with 1× PBS. Stained plates were scanned using Epson scanner V370. To estimate the amount of colony formation, crystal violet was dissolved in 100% methanol. In a 96-well plate, the crystal violet was diluted in 1:10 and absorbance was measured at 540 nm using Tecan Spark 10 M plate reader instrument.
Spark 10 m plate reader instrument
Manufactured by Tecan
The Spark 10 M is a multimode plate reader instrument designed for a wide range of microplate-based applications. It offers high-performance detection of absorbance, fluorescence, and luminescence signals.
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Lab products found in correlation
2 protocols using spark 10 m plate reader instrument
1
Quantitative Evaluation of Colony Formation
2
Functional Assay for CEBPB-AS1 Knockdown
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One thousand cells per well plated onto six-well plates were transfected with either control siRNA or siRNA against CEBPB-AS1 for 48 h and then retransfected for another 48 h. Colonies were grown for 4–5 days, with media change every 2 days. Cells were fixed using 4% buffered formaldehyde, stained with 0.05% crystal violet, and plates were scanned using Epson scanner V370. To estimate number of colonies, crystal violet staining was dissolved in 100% methanol, diluted to 1:10 in PBS and absorbance was measured at 540 nM using Tecan Spark 10M plate reader instrument.
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