Iam facing a problem with stable clone.I think Iam having a problem but not so sure. I have c3h10 cells which is a stable clone with G418 resistance and the stably expressing protein is a cell cycle probe for G1 (red tag). In this I have to insert a protein (green tag) which is localized to the nucleus, which has puromycin resistance. I followed the standard procedures,selected using G418 and puromycin, and i looked for the green and red fluorescence. What I saw was that, not all cells were expressing both the tags. There were some cells which didnt express both.
So, I assumed that the cells not expressing both the tags may be in the G2 /S or M phase. is this correct or is something really wrong with my expt. please help.