DNA Station
Treatment of Cells (WEHI 231 cells) with anti-IgM in order to examine proliferation and transcription of early response genes in B cells in which the B cell receptor has become activated.
All following steps are required to be carried out in a tissue culture laminar flow fume hood to provide for aseptic technique.
1. COunt cells, Add 10 to the 6 cells to a round bottom tube. Make up a volume of around 0.5 mL with RPMI medium, which must be supplemented with 10% FCS and antibiotics to prevent contamination.
2. Add 0.5 mL of 10 ug/mL goat anti-IgM to the cells.
3. A negative control will be made through adding 0.5 mL medium.
4. Incubate cells in a 37 celsius waterbath for 45 minutes and then purify RNA.