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Each sample run was recalibrated by infusion with the same TFA-NA calibrant before the chromatographic front.
The sample run was considered acceptable if the ratio to the control probe pairs was between 0.8 and 1.2.
The amount (in pM) of library sample run was determined by quantitative PCR so that the same cluster density was present in each lane.
The lymph node tissue designated for OSNA (maximum weight of 600 mg per sample run) was shock frozen and stored at −80 °C until further use.
Spontaneous and rewarded alternation was assessed as previously described (Bannerman et al., 2004), except for rewarded alternation where a free choice on the sample run was permitted.
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The apparatus and a sample run are depicted in supplementary material (Methods S1).
DTASelect-filtered data for each sample run were then prepared for semi-quantitative analysis.
Two sample runs were included to demonstrate the concept of dynamic causal network modification and time-delay management.
Agreement of HPLC data with analyzed standards prior to sample analyses and in between ten sample runs were satisfactory.
Sample runs were monitored using the SprayQC quality monitoring system (Scheltema and Mann 2012).
For each sample, triplicate runs were performed as independent experiments.
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