I am trying to grow up monoclonal HEK cells by sorting single cells into 96-well or 384-well plate. However, in the majority of wells the cells simply did not proliferate. In 96-well plate, I could get about 20 monoclonal lines. In 384-well plate the was even lower. I am thinking of using live/dead cell staining to exclude dead cells. Does anyone know other tricks that may help to increase the number of monoclonal lines?
Actually 20/96 is not that bad. It depends on what you need them for. If it is a stable transformation, you may slightly increase your original seeding density, i.e.2-3 fold. You have to be careful though as too many cells seeded will give you polyclonal population, which requires recloning down the road. Also, check the viability of your cells with Trypan Blue - maybe you have too many dead cells in your original population.
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