GSM8508589: Alzheimer's disease sample 11; Homo sapiens; RNA-Seq
Total RNA including miRNA was extracted from the synaptosomes of AD and control postmortem brain samples usingthe TriZol reagent with some modifications. Poly(A) RNA sequencing library was prepared following Illumina's TruSeq-stranded-mRNA sample preparation protocol. RNA integrity was checked with Agilent Technologies 2100 Bioanalyzer. Poly(A) tail-containing mRNAs were purified using oligo-(dT) magnetic beads with two rounds of purification. After purification, poly(A) RNA was fragmented using divalent cation buffer in elevated temperature. The DNA library construction is shown in the following workflow. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-ended sequencing was performed on Illumina's NovaSeq 6000 sequencing system. All extracted RNA was used in the library preparation following Illumina's TruSeq -small-RNA-sample preparation protocols (Illumina, San Diego, CA, USA). Quality control analysis and quantification of the DNA library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Single-end sequencing 50bp was performed on Illumina's Hiseq 2500 sequencing system following the manufacture's recommended protocols