...kinase promoter drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO...sulfate, and puromycin are located in the “Appendix”. F.2. Determining the Optimal Puromycin Concentration...differently to puromycin selection. Addgene strongly recommends that you determine the optimal puromycin concentration...to fresh puromycin-containing media every other day. f. The minimum concentration of puromycin that results...with finer increments of puromycin to determine a more precise optimal puromycin concentration.) F.3. Protocol...expression of the puromycin resistance gene. f. To select for infected cells, add puromycin to the media at...positive control for the puromycin selection. Days 4+: g. Change to fresh puromycin-containing media as needed...

...Chloramphenicol 25 µg/mL Coumermycin 25 µg/mL Gentamycin 10 µg/mL Kanamycin 50 µg/mL Spectinomycin 50 µg/mL Tetracycline...

...obtaining accurate titers. Check the cells for mycoplasma regularly Do not over or under-grow your cells... after 20-30 passages. Do not add penicillin/streptomycin to the media. Titer will vary between cell lines... . The following day, cells were treated with puromycin. Colonies were allowed to grow in the presence...

...deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin), which enables selection of a stable...preparation Appropriate antibiotic for selection (e.g. puromycin, blasticidin) Reagent Preparation DMEM Complete...obtaining accurate results. Check the cells for mycoplasma (Link opens in a new window) regularly Do not... after 20-30 passages. Do not add penicillin/streptomycin to the media. Infection efficiency will vary...

... µg/mL Coumermycin 25 mg/mL (dissolve in DMSO) 25 µg/mL Gentamycin 10 mg/mL 10 µg/mL Kanamycin 50 mg/mL.../mL 50 µg/mL Spectinomycin 50 mg/mL 50 µg/mL Tetracycline 10 mg/mL 10 µg/mL Note: Unless otherwise indicated...

...obtaining accurate titers. Check the cells for mycoplasma (Link opens in a new window) regularly Do not... after 20–30 passages. Do not add penicillin/streptomycin to the media. Titer will vary between cell lines...