HeLa cells freezing protocol

Authors

Heike Stichnoth

Materials

Medium

1. 450 ml DMEM (GIBCO 1.0 g/l Glucose; Cat. # 31885-023)

2. 50 ml FBS (10% GIBCO Cat. # 10270-106)

3. 1 ml Primocin (Invivogen 50mg/ml Cat. # ant-pm-1)

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

Procedure

1. trypsinize 10 flasks with 2ml Tryp. 0.25% for ~ 10 min / 37ºC

2. suspend cells in some medium (~ 8ml for 3 flasks)

3. pool the cell suspensions in a 50ml centrifuge tube

4. centrifuge 5min/1500 rpm

5. remove the supernantant

6. resuspend the cell pellet in 20ml freezing medium (regular growth medium + 5% sterile DMSO)

7. aliquot in 20 x 1ml Cryo tubes

8. freeze over night at -80 °C (in a box with numbers on the lid)

9. then transfer the tubes in the liquid N2 - tank

Further details

Protocol provided by Heike Stichnoth.

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• Monday, 02 June 2014 - 14:26 UTC

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