User:Elsealab/sandbox

Thawing Cells 1.) Look up cells in the Freezer Log and record the coordinates. 2.) Mark the Freezer Log with the date and your initials 3.) Remove cells from freezer location (-80 or LN) 4.) Allow cells to thaw at room temperature 5.) While the cells are thawing, label your flasks or plates and conicals 6.) Pipet cells from nunc tubes into a labeled conical 7.) Spin cells at 1,200 rpm for 10 minutes at 4C 8.) Remove the supernatant and leave the pellet 9.) Add the appropriate amount of media to reconstitute cells 10.) Pipet up and down to throughly mix cells so there is no clumps 11.) Pipet cells into flask or plate 12.) The next day remove media and add fresh media.

Tips: Some cells grow slower than others and very few cells acutally survive the freeze thaw process. Try thawing and combinting two tubes of cells to get better growth. Sometimes cells can take longer than 12 -18 hours to adhere. Try supplementing your media with 20% FBS instead of 10% and get your cells to adhere and grow.
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