Difference between revisions of "Freezing of A1 cells"
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(Created page with "''in one 175 cm2 flask'' Materials: - APBS (100 ml PBS + 25 ml diwater) - TE in APBS (36 ml APBS + 4 ml TE) - 1x 15 ml FALCON tubes - Pipettes - 1 cryo tube - freezing media ...") |
(No difference)
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Revision as of 08:30, 12 January 2015
in one 175 cm2 flask
Materials: - APBS (100 ml PBS + 25 ml diwater) - TE in APBS (36 ml APBS + 4 ml TE) - 1x 15 ml FALCON tubes - Pipettes - 1 cryo tube - freezing media
Procedure:
- aspirate the old media from the cells - wash the cells 1 times with 5 ml APBS - remove the APBS - add 5 ml TE - wait till the cells are detached - stop the TE with HS – media - centrifuge at 1000 rpm, 3 min at 4ºC - aspirate the supernatant - dissolve the pellet into 1 ml freezing media - transfer the cells into the cryo tube and label the tube with the cell name, passage number and date - store the tube for one day at –80ºC - store the vial in liquid N2