i. Dilute antibiotic in culture media to a dose of 10 mg/mL. Dilute 1:1 in culture media four times to yield a two-fold dose response ranging from 625 ng/mL to 10 mg/mL. ii. Aspirate media from cultured cells and replace with antibiotic selection media. Replace normal media in remaining well for an untreated control. iii. Monitor cells daily with a standard light microscope or by imaging using an Incucyte. For most cell lines, puromycin selection will be complete within 48 h. iv. Determine the minimum antibiotic dose at which selection is complete in uninfected cells ( Figure 1B). If death is not apparent from morphological features of the selected cells, SYTOX Green may be used, as described below, to confirm cell death. v. Note the minimum dose required to kill >99% of uninfected cells and use this dose for selection of NucLight Red-infected cell lines in step 3.

### Language ###
English

### Assessment ###
The text contains specific timing information about puromycin selection, which could be a natural point of inquiry for lab researchers.

### Query ###
Tell me the typical duration needed for puromycin selection to complete.
Tell me the typical duration needed for puromycin selection to complete.