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They still eat pellets derived from wild fish, but the fish in the pellets comes from discarded cuts of wild fish that are said to be caught in a sustainable manner.
The rinse pellets derived from the foregoing samples were also subjected to duplex PCR in parallel.
RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion.
All samples containing abscess material, as well as the respective pellets derived from liquid cultures, were extracted in the same experiment in accordance with the kit manufacturer's instructions.
The final analysis was then performed as follows: first, the rinse pellets derived from randomly selected 100 samples that yielded germ tube positive yeasts were subjected to duplex PCR.
Briefly, the cell pellets derived from cells grown to the mid-logarithmic phase were resuspended in ice-cold thylakoid buffer (50 mM 3- N-morpholino -propanesulfonic acid, pH 7.0, 0.4 M sucrose, 5 mM MgCl2, 5 mM CaCl2, 1 mM freshly made phenylmethylsulfonyl fluoride).
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RNA was extracted from mycelium pellets deriving from two independent culture samples using the Column Plant RNAout (TIANDZ) according to the standard protocols.
The pellet derived from the centrifugation step was used for the determination of starch.
The cell pellet derived from two 60-mm dishes, cultured as described above, was resuspended in 1 ml of Trizol (Invitrogen, Carlsbad, CA, USA).
As determined by densitometric analysis, Cytochorme C decreased by 35% only in the mitochondrial pellet derived from cells expressing both Bcl-2 and mutSOD1 (Fig. 2C-right panel).
Amplification and direct Sanger sequencing of the sigma C protein gene from the cell pellet derived from the fourth tissue culture passage demonstrated identical sequence to that derived from the post-mortem tissue samples.
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