HeLa cells thawing protocol

Authors

Heike Stichnoth

Materials

Medium

• 450 ml DMEM (GIBCO 1.0 g/l Glucose; Cat.# 31885-023)
• 50 ml FBS (10% GIBCO Cat. # 10270-106)
• 1 ml Primocin (Invivogen 50mg/ml Cat. # ant-pm-1)

... sterile filtered through 0.22 μm Milli Pore Bottle Top Filter

Procedure

• suspend cells in 4ml mediu$m and transfer into 15 ml centrifuge tube
• centrifuge 5 min/1500rpm
• aspirate supernantant
• resuspend in 3ml medium
• seed out into a T75 flask, provided with 15 ml medium

Further details

Protocol provided by Heike Stichnoth.

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Comments

• Mateusz Putyrski Sunday, 10 March 2013 - 23:25 UTC

  1500 rpm is not really informative
  for me 3 minutes at 800x g work pretty well
  and 10 mL medium in a T75 flask is surely enough