GSM9967068: Diabetes mellitus and periodontitis mouse brain, pooled RNA; Mus musculus; RNA-Seq
Brain samples from six mice per experimental group were pooled prior to library preparation. Total RNA, including miRNAs, was extracted from brain samples using the mirVana miRNA isolation kit (Thermo Fisher Scientific, Waltham, MA, USA). Extracted total RNA was quantified using a NanoDrop One spectrophotometer (Thermo Fisher Scientific). RNA quality was evaluated using an Agilent 2100 Bioanalyzer with the Agilent RNA 6000 Nano kit (Agilent Technologies, Santa Clara, CA, USA). Total RNA obtained from each sample was used for sequencing library construc-tion using NEBNext Ultra II Directional RNA Library prep for Illumina (New England Biolabs, Ipswich, MA, USA) and NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), with NEBNext Multiplex Oligos for Illumina (96 Unique Dual Index Primer Pairs) (New England Biolabs). Library quality was assessed using an Ag-ilent 2200 TapeStation (Agilent Technologies, Inc.) and Applied Biosystems 7500 Re-al-Time PCR system (Thermo Fisher Scientific, Inc.) with High Sensitivity D1000 rea-gents (Agilent Technologies, Inc.) and a GenNext NGS Library Quantification kit (Toyobo Co., Ltd., Osaka, Japan). The pooled libraries generated from the samples were sequenced using Illumina NextSeq 500 (Illumina, Inc., San Diego, CA, USA) with 75-base-pair (bp) single-end reads.