Hello all. We have created GFP macrophages using the pIRESneo vector with the EGFP gene inserted into the MCS. The cells were then selected using G418. Here's the problem...even with selection we have two distinct populations, one negative and one positive. We are now trying to sort the cells, but cannot eliminate the negative population. The best sort results that we have accomplished is 60% positive and 40% negative populations. Does anyone have any clues or suggestions on why this is happening and what we can do to reduce/eliminate the negative population? We had hopes of using these cells for in vivo studies. Thanks, Marcia Marcia Hart GRA Division of Biology Kansas State University 19 Ackert Hall Manhattan, KS Office:(785)532-6795 E-mail: mlh2207@ksu.edu
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