Was hoping you might help me solve an issue i'm stumped on. I'm using phoenix cells to produce a virus to infect HT1080 cells with a gene of interest. The virus contains puro-resistance as well as a GFP marker tagged to a gene of interest. After infection and subsequent selection, many cells died while a few lived (as expected). When the plates were grown up with only resistant cells, not all of the cells expressed GFP. In fact, only about 20% i'd say of a 10mm conf. dish were GFP positive. I've done this with 3 different plates infected with 3 GFP-gene of interest viruses. I am stumped because even if you had a super rare event where a cell only obtained Puro R, you wouldn't expect to see it on multiple plates.
Puro Selection Issue
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