... confluent, expand into larger vessels. A confluent well of a 6-well dish can be expanded into a 10 cm...reagent Day 3-14: Change media as needed Day 14-18: Expand and harvest stable cell lines Equipment Biosafety...undergo additional seleciton while the population expands. At this time, some researchers reduce the concentration...cm dish. A confluent 10 cm dish can be expanded into two 75 cm 2 flasks. *Pro-Tip* This selection method...populations are growing well and have been sufficiently expanded, prepare cell stocks and/or harvest to test for...will have higher levels of expression. Consider expanding populations transduced with a variety of dilutions...Figure 1: Generation of monoclonal cell lines from expansion of individual A549 cells stably expressing Cas9...

... Monitor cells for growth and expand cells Day 14–30: Analyze and expand monoclonal lines of interest ...cells have expanded but before they become over-confluent, trypsinize the cells and expand them to larger...After the monoclonal lines have been sufficiently expanded, screen the lines for transgene expression and...Figure 1: Generation of monoclonal cell lines from expansion of individual A549 cells stably expressing Cas9... in individual wells of a 96-well plate after expansion of single cells for 13 days. Note that growth ...Single cells from a stable cell pool were then expanded for an additional 3 weeks under blasticidin selection...stable cell pool (pool) and from individual cell expansions (monoclonal, 1-5). Wild-type A549 cells do not...

...limited MCS (BamHI - EcoRI - SalI) and you want to expand that with the addition of NdeI, PacI, AscI, and...

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