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I can't find the LG2 cell line that you mentioned but often times you can find karyotype information on ATCC or COSMIC (for cancer cells)


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Might be considered "not answering the question" but: Option 1: Depending on the type of cells you are using, you can use milder methods to detach cells. Example include Versene which is essentially PBS + EDTA. Milder still is to just use PBS that has no Mg or Ca (no chelation). Option 2: Duplicate your setup - instead of two dishes - one with drug, one ...


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It would be stressful for cells to be trypsinized 24 hours after seeding. After cells are plated, there is a lag time to start growing. Perhaps, during the lag time, physiological state of cells is stabilized. They have to express proteins digested by trypsin and display on the surface etc. Then you could disturb the state again by trypsinizing cells again ...


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It is not advisable to trypsinize the cells too often when you have to maintain them. However, in your case the situation is different. Time required to attach depends on cell type. In any case while performing an experiment, your cells should not be already under stress. Therefore you should seed the flasks/culture plates with a smaller concentration of ...


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It's because E.coli BL21(DE3) are competent cells. The competent is the key here as the cells were chemically treated so the transfection can be performed by heat-shock with high efficiency. This means these bacteria are quite fragile, due to the chemical treatment, and therefore are very sensitive to both mechanical and thermal shocks. Pre-heating the ...


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Although I have not yet done, in general, cells are happier in a conditioning medium. Grow the original cells in 10cm dishes and save the medium and use it to grow single colonies. You might want to mix used and fresh medium (1:1 ratio). Growth factors are another option. Insulin is sometimes used, but it also depends on cells. Coating the surface of well ...


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This should be relatively straight forward. Sort the cells into single wells so all descendants are clones of the sorted cell and let them grow until they are dense enough to be transferred into a bigger plate (probably 48 or 24 well). Continue this process until you can go into big cell culture flasks or plates and make sure that you freeze away a number of ...


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TGF-beta would be a good candidate. To cite: "TGF-β inhibits G1/S progression in a variety of eukaryotic cell types. Among these, untransformed epithelial cells are particularly sensitive to the growth inhibition by TGF-β." http://genesdev.cshlp.org/content/14/24/3093.full Fetal bovine serum (FBS) contains a high level of latent TGF-β. Human serum as well ...



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