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After incubation, 2 l of 5 First-strand RT buffer, 0.5 l of 0.1 M dithiothreitol (DTT), 0.5 l of RNaseOUT (40 U/l), and 0.5 l SuperScript III Reverse Transcriptase (200 U/l) were added and incubated at 25 C for 10 min, and then 50 C for 50 min, followed by the final termination at 85 C for 5 min

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Adequate caloric intake is essential in amyotrophic lateral sclerosis and future studies should monitor malnutrition closely (168)

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