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Organs from five rats were pooled to account for biological variation.
Therefore, data from A C and D E trimmed rats were pooled.
Eight CPs from four rats were pooled for each sample used downstream for PCR, and two CPs from one rat were used for each IHC sample.
Briefly, fresh forebrains of four rats were pooled, gently homogenized in 10 volumes (v/w) of ice-cold buffer (5 mM Tris-HCl pH 7.4, 50 μM CaCl2, 0.5 mM dithiothreitol), and supplemented with 10% sucrose.
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An equal amount of the total cDNA from each of the three rats was pooled, and the pooled solution was used as the template material for RT-qPCR.
RNA from three rats was pooled for each array.
For the sample stability test, urine from natrosol-, furoremide-, or HCBD-treated rats was pooled treatment-wise (5 males and 5 females per treatment group).
Hypothalami of three male Sprague-Dawley rats (supplier Harlan, Netherlands) were pooled and the frozen tissue was homogenized in 0.5 ml of low salt buffer (10 mM Hepes, pH 7.9, 1.5 mM MgCl2, 50 mM KCl, 0.5 mM dithiothreitol, proteinase inhibitors) using a rotor-stator homogenizer (ART Labortechnik, Müllheim, Germany).
Calvaria from eight rats per sex were pooled for each experiment.
The samples from incubation with rat liver microsomes were pooled into designed cassette groups and analyzed by HPLC/electrospray ITMS in full-scan mode.
Rat tissue lysates were pooled in order to handle a large number of samples that exceeded the lane capacity of a gel, while human homogenates were not pooled.
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