RNA-Sequencing of EC and PVC from aged APOE mice

RNA-Sequencing of entorhinal cortex and primary visual cortex from 14-15 month old APOE3/4 vs. APOE3/3 targeted replacement mice. Entorhinal cortex and primary visual cortex tissue from 14-15 month old APOE mice (10 male APOE3/3s and 19 male APOE3/4s) were collected, and RNA was extracted using Trizol. Starting with 2 ug of total RNA per sample, Poly(A)+ mRNA was purified, fragmented and then converted into cDNA using the TruSeq RNA Sample Prep Kit v2. For RNA-Sequencing of the cDNA, we hybridized 5 pM of each library to a flow cell, with a single lane for each sample, and we used an Illumina cluster station for cluster generation. We then generated 149bp single end sequences using an Illumina Hi-Seq sequencer.针对14-15月龄APOE3/4与APOE3/3靶向替换小鼠的内嗅皮层（entorhinal cortex）及初级视觉皮层（primary visual cortex）开展RNA测序（RNA-Sequencing）。我们收集了14-15月龄APOE小鼠的上述皮层组织，其中包含10只雄性APOE3/3小鼠与19只雄性APOE3/4小鼠，并采用Trizol试剂提取总RNA。每份样本取2 μg总RNA，经Poly(A)+富集纯化得到信使RNA（mRNA），随后进行片段化处理，并使用TruSeq RNA样本制备试剂盒v2（TruSeq RNA Sample Prep Kit v2）反转录为互补DNA（cDNA）。在对上述cDNA进行RNA测序时，我们将每个文库的5 pM样本与流动槽（flow cell）进行杂交，每个样本单独占用一个泳道，并使用Illumina集群工作站完成集群生成。随后依托Illumina Hi-Seq测序仪生成149 bp单端测序序列。