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The brains were placed in 30 % sucrose and 0.05 % sodium azide solution overnight at 4 °C.
In brief, the animals were decapitated and brains were placed in cold sucrose artificial cerebrospinal fluid (sucrose-ASCF, in mmol/L: sucrose 213, KCl 2.5, NaH2PO4 1.25, NaHCO3 26, D-glucose 10, MgSO4 2, CaCl2 2, pH 7.4, 0 4 °C, saturated with 95% O2 and 5% CO2).
Subsequently, the brains were placed in 30% sucrose solution for 48h before sectioning.
For in vivo ChIPs, P0 brains were placed in 1% formaldehyde immediately after dissection.
Twenty h later brains were placed in 70% ethanol until they were included in paraffin.
Brains were placed in the IVIS chamber and background fluorescence was set as zero.
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The left halves of the brain were placed directly into 2 mL of isotonic saline with 0.167% gelatin (gel saline).
The brain was placed in an ice-cold isolation media containing 0.25 M sucrose, 0.5 mM EDTA (dipotassium salt) and Tris HCl (pH 7.4) immediately after the removal.
The whole brain was placed in a hexagonal polystyrene weighing boat (top edge side length, 4.5 cm; bottom edge side length, 3 cm) and was covered with powdered dry ice.
After every two hours of work Gordon's brain is placed back in the incubator, where it remains active even though it's not controlling a body, a state Warwick equates with our experience of dreaming.
Half of each bisected brain was placed in formalin and subsequently embedded in paraffin.
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