View source for XL2013-cell-colony-formation-assay

==Single cell colony formation assay #48 well plate==
# 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)
# 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).
# Wells were examined microscopically and those containing only a single clone were selected for analysis.
# Stained with 0.5% crystal violet:
## remove the medium
## wash with PBS once
## add ethonal for 10min with lid on
## open the lid and dry in the air completely
## add 500 ul 0.5% crystal violet for 15 min
## wash with water and dry it
# Count the colony and calculate the colony formation efficiency:
 no. of colony /48 *100%

==Colony formation assay #6 well plate==
# 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
# Wells were examined microscopically after 7-10 days,
# Stained with 0.5% crystal violet:
## remove the medium
## wash with PBS once
## add ethonal for 10min with lid on
## open the lid and dry in the air completely
## add 500 ul 0.5% crystal violet for 15 min
## wash with water and dry it
# Count the colony and calculate the colony formation efficiency:
 no. of colony /500 *100%
Xiao 2013, Bergen

[[Category:Protocols]]
[[Category:Xiao Liang]]
[[Category: Bergen]]
[[Category: Norway]]
[[Category: 2013]]