Writing down points Arman showed me on 20191130

Dan Yamamoto-Evans 2019/11/30 14:17

Material

  • FBS
  • DMEM medium
  • x100 Penicillin-Streptoadivin
  • Cell stock




Protocol

1 Preparation
  • Warm the FBS (50mL, stored at -20˚C) and DMEM (500mL, stored at 4˚C) in the water bath @37˚C.
  • Thaw x100 Pen-strep
  • Add 50 mL of FBS to DMEM (Final 10%)
  • Add 5 mL of x100 Pen-strep to DMEM (Final x1)
  • Label the bottle, mix and warm in the water bath.




2 Cell stock
  • Take out the cell-stock from liquid nitrogen tank, and put on ice.





  • Aliquot 5 mL of the warm media to a new falcon tube.
  • Thaw the cell-tube on the water bath, and once it is thawed, add the entire sample to the aliquotted medium.




3 Seading
  • Spin down sample at 1000 rpm for 5 min.
  • Remove sup.
  • Add 10 mL of the same medium to the tube, and resuspend by pippette to mix.
  • Take out an 10cm dish, and label it.







  • Add the sample to dish, and spread cells by moving plate horizontally and than vertically.
  • Incubate in the human cell culture incubator.
Tips
  • Clean the spcace with EtOH before/after use.
  • DO NOT put lid from reagents etc on the surfaces.
  • EtOH stuff, but NOT pippettes and plates.
  • Put back the medium in Human medium fridge.

Dan Yamamoto-Evans 2019/11/30 15:54

  • interactome/cell_cultivation1.txt
  • Last modified: 2019/12/13 19:00
  • by dan