XL2013-cell-colony-formation-assay

Single cell colony formation assay #48 well plate

1. Typsinize the cells into single cell suspension and put 2ml of cell suspension into 35mm dish (cell density can be adjusted to make it easy to collect the single cells)
2. Pick single cells by using 10ul pipette (2ul); or cells were seeded directly into 96-well plates by using the single cell plate sorting function of the FACS Aria sorter (BD biosciences).
3. Wells were examined microscopically and those containing only a single clone were selected for analysis.
4. Stained with 0.5% crystal violet:
   1. remove the medium
   2. wash with PBS once
   3. add ethonal for 10min with lid on
   4. open the lid and dry in the air completely
   5. add 500 ul 0.5% crystal violet for 15 min
   6. wash with water and dry it
5. Count the colony and calculate the colony formation efficiency:

no. of colony /48 *100%

Colony formation assay #6 well plate

1. Typsinize the cells into single cell suspension and put a total of 500 cells/well were seeded in 6-well plates within 3 mL complete culture medium in 6 well plate
2. Wells were examined microscopically after 7-10 days,
3. Stained with 0.5% crystal violet:
   1. remove the medium
   2. wash with PBS once
   3. add ethonal for 10min with lid on
   4. open the lid and dry in the air completely
   5. add 500 ul 0.5% crystal violet for 15 min
   6. wash with water and dry it
4. Count the colony and calculate the colony formation efficiency:

no. of colony /500 *100%

Xiao 2013, Bergen