GSM9241464: MeHg_2; Mus musculus; RNA-Seq
Frozen brain tissue samples were each suspended in 1 mL of TRIzol (cat#15596026, Thermo Fisher) reagent and were added directly to a microcentrifuge tube. The tissue was homogenized using a tissue homogenizer and incubated for 5 minutes at room temperature. 200 μL of chloroform (cat#C2432-1L, Sigma-Aldrich) was added to the mixture, which was then vortexed for 15 seconds to ensure thorough mixing. The samples were incubated at room temperature for 2-3 minutes and then centrifuged at 12,000 x g for 15 minutes at 4°C. The aqueous phase was carefully transferred to a new RNAse-free tube, and an equal volume of 70% ethanol was added. Washes were performed using the PureLink RNA Mini Kit (cat# 12183020, Thermo Fisher). The extracted RNA was stored at -80°C for long-term preservation. The quantity and purity of the extracted RNA were assessed using a spectrophotometer. RNA samples from biological triplicates (both female and male mice) were isolated as previously described and sent to Sacramento, California, for RNA-Seq analysis at Novogene Bioinformation Technology Co., Ltd. (Sacramento, USA). Sequencing was performed using the Illumina NovaSeq platform with paired-end 150 base pair reads (PE150) and reference genome mm39.