Illumina HiSeq 4000 paired end sequencing; RNA-seq of mouse brain tissue to investigate the integrity of the blood brain barrier to trimethylamine N-oxide
RNA-seq of mouse brain tissue to investigate the integrity of the blood brain barrier to trimethylamine N-oxide
Mice were transcardially perfused with 0.9% saline at 4 °C to remove circulating blood, and brains were removed and collected into RNAlater (Thermofisher Scientific Ltd., UK) prior to storage at -20 °C for later analysis. TMAO-treated mice were injected i.p. with 1.8 mg/kg TMAO in 100 microlitres saline vehicle, a dose calculated to approximate human circulating TMAO levels. Saline-treated animals received only 100 microlitres saline. Whole brain total RNA was extracted using a PureLink RNA Mini Kit (Thermofisher Scientific Ltd., UK). TruSeq Stranded mRNA LT Sample Prep Kit as per the manufacturer's instructions.