The cells and medium were pooled and titered on PK15 cells.
For the purification of the fusion proteins IFP-6xHis and TPK1-TEV-IFP-6xHis, L. tarentolae cells from nine 150 cm2-tissue culture flasks each containing 60 mL of non-selective expression medium were pooled and centrifuged.
Colonies that could grow in minimal medium were pooled together and used for screening of PRA isomerase activity in the E. coli trpF − strain FBG-Wf, as before.
Total RNA extracted from 37 h old cultures of S. rolfsii grown in EPSmin17 and EPSmax13 medium were pooled in a 1 1 ratio to guarantee equal predominance of both RNA populations and subsequently reversed transcribed into cDNA.
We also found some overlap with prior microarray experiments in which we compared gene expression in vegetative and sexual mycelia; however, in these experiments we used mycelia grown in defined medium [ 9], whereas for RNA-seq analysis, RNA from mycelia grown in defined medium and cornmeal medium were pooled.
The remaining cells, which had been isolated from two fetuses, were resuspended in 200 μL William's medium with or without 25 μM BPA-GA and incubated at 37°C for 10, 30, 60, or 120 min. At each time point, cells and medium were pooled and extracts were prepared for HPLC as described above, by adding the same volume of methanol.
Medium from 20 drops was pooled and centrifuged at 14,000×g for 10 minutes at 4°C.
Mouse epiphyseal chondrocytes were seeded at 5×10 cells per well and the medium from 6 wells was pooled for analysis.
Adult pig articular chondrocytes, which are significantly smaller than the mouse epiphyseal cells, were seeded at 3×10 cells per well and the medium from 6 wells was pooled for analysis.
Conditioned medium from three independent experiments was pooled, aliquoted and stored at -20 °C until required.
