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All the dCas9-expressing transformants were maintained with 60 µg/ml G418 for a few days before imaging.
Transformants were maintained in HL5 at 20 µg/ml G418, 50 µg/ml hygromycin B or 10 µg/ml blasticidin S, as appropriate.
The transformants were maintained on MM plates (1% glucose, 10 mM NH4Cl, 10 mM potassium phosphate (pH 6.5), 7 mM KCl, 2 mM MgSO4).
Transformants were maintained on minimal medium agar (Samson et al. 2004) supplemented with 100 µg/ml Hygromycin B. Alkane production was studied on glucose (composition same as minimal medium), oatmeal and Yeast Malt medium (YM, 3% yeast extract, 3% malt extract, 10% glucose).
Transformants were maintained under 2.5 μg/ml neomycin and 2.5 μg/ml hygromycin selection, and RNAi was induced by adding 1 μg/ml tetracycline.
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ET12567/pUZ8002 transformants was maintained in chloramphenicol (25 μg/ml), kanamycin (25 μg/ml) and apramycin (50 μg/ml).
A. nidulans FAC transformants (Table 1) were maintained on culture plates for three generations for phenotype and chemical screening.
To obtain stable transformants, the LLC cells were maintained in DMEM containing 10% FCS and 1200 μg/mL hygromycin B (Wako, Tokyo, Japan).
Transformants of A. cellulolyticus YP-4 were maintained on MM agar plates (Fujii et al. [2012]).
Transformants were subsequently maintained in 1 μM triclosan (LBT).
These results indicate that ura- selection is required for the transformant to show laccase activity, which is maintained in ura- selective cultured media.
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