Wittrup: Thawing Protocol

Materials

warm recovery media

Protocol

Quickly warm a vial of frozen cells by immersing in a 37 degree water bath.

Add 2 ml recovery media and centrifuge for 10 min at 200 x g (low speed).

Discard supernatant.

Resuspend cells in 1 ml and plate out in 9 ml recovery media on a 10 cm dish.

Check for adherance the following day.

Feed and subculture as necessary.