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LIQUID CULTURE OF WORMS WITH FERMENTOR

. The worm pellet should be clean and light brown. Try to remove as much of the supernatant as you can. About 20 ml worm pellet should be collected by this method from the 2-liter culture in the batch fermentor. Freeze the tubes, containing 1 ml 0.1 M NaCl

OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS

into a 60mm culture dish containing 5m1s selection medium. 10. When the clone reaches 30% confluence, passage into 3 x l0()mm culture dishes containing l()mls selection medium per dish. 3 dishes are used so that two can be used for RNA isolation

Purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion

on ice for 30 min (all antibodies 1/100 v/v). 4.  Wash with 10 mL of PBS/FBS, spin and resuspend in PBS/FBS with 1 µg/mL of propidium iodide. Filter through 70 µm nylon mesh. 5. Select cells which are PI negative, c-kit positive, Sca-1 positive