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Inocula for batch experiments in flasks were prepared by picking a single colony of C. necator H16 from a streaked plate and inoculating Luria Bertini (LB) broth in glass tubes.
The construct purity was determined by picking a single bacteria colony to PCR reaction mixture in the conditions described above.
The bottlenecks were accomplished by picking a single colony and streaking for single colonies approximately every 2 d; the process was repeated eight times.
The isolate was purified by picking a single uredinium using a fine needle under an Olympus SZX12 dissecting microscope and reinoculating it on leaves of Williams 82.
Clonal cells expressing either GFP-TRF1 or GFP-TRF2 from each cancer cell line were generated by picking a single cell colony.
For each overnight culture, 5 mL of tryptone broth (10 g/L tryptone and 10 g/L NaCl) was inoculated by picking a single colony and incubated in a 37 °C shaker for 16 18 h.
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