Sorted cells_PS2APP brains_7/13mo

Mice of indicated ages and genotypes were perfused and their brains dissected and dissociated. Cells were fixed, immunolabeled and FACS sorted. RNA was extracted from neuron, astrocyte, and microglial cell populations. Typical RIN=4-5 for neurons, 6-8 for astrocytes, and 5-7 for microglia. Typical RNA yields ~100ng for neurons, ~20ng for microglia, and ~10ng for astrocytes. cDNA was generated from up to 25 ng of total RNA using Nugen's RNA-Seq method for low-input RNA samples, Ovation RNA-Seq System V2 (NuGEN). (Per manufacturers instructions, total RNA was neither depleted of rRNA nor polyA-selected.) 1 ug of sheared cDNA was taken into further processing, starting at end repair step, using Illumina's TruSeq RNA Sample Preparation Kit v2 (Illumina). The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0006149 Astrocytes, microglia and neurons were sorted from 7- or 13-month old PS2APP or non-transgenic mice, 4 <= n <= 7 per group.对指定年龄和基因型的小鼠实施灌流处理，随后解剖并解离其脑组织。将所得细胞进行固定、免疫标记及荧光激活细胞分选（FACS）。分别从神经元、星形胶质细胞和小胶质细胞群中提取RNA。各类细胞的典型RNA完整性数（RIN）为：神经元4~5，星形胶质细胞6~8，小胶质细胞5~7。典型RNA产出量为：神经元约100ng，小胶质细胞约20ng，星形胶质细胞约10ng。使用纽根（Nugen）公司针对低起始量RNA样本开发的Ovation RNA测序系统V2（NuGEN），从最多25ng的总RNA中合成cDNA。（按照制造商操作说明，总RNA既未去除核糖体RNA（rRNA），也未进行polyA分选。）取1μg剪切后的cDNA，以Illumina公司的TruSeq RNA样本制备试剂盒v2（Illumina）为工具，从末端修复步骤开始开展后续处理。本数据集的"SAMPLE_ID"样本特征为基因泰克（Genentech）内部使用的样本标识符。该项目在基因泰克的ExpressionPlot数据库中的编号为PRJ0006149。本实验从7月龄或13月龄的PS2APP转基因小鼠及非转基因小鼠中分选星形胶质细胞、小胶质细胞与神经元，每组样本量为4≤n≤7。