RNA pol II ChIP-seq LR14 and HFF human fibroblasts

The aim of the experiment is to determine if RNA pol II occupancy and abundance is altered human fibroblasts from a patient that has a mutation in the gene Taf8, which plays an important role in RNA pol II mediated transcription. Overall design: I performed ChIP with an antibody against RNA pol II, and have submitted the precipitated DNA from patient and control samples (L-IP and H-IP, respectively). In order to aid with normalisation, I spiked the fibroblast chromatin with 1/10 drosophila chromatin. Input for the HFF sample will also be submitted.

本实验旨在探究携带TAF8基因突变的患者来源的人成纤维细胞中，RNA聚合酶II（RNA pol II）的结合占有率与分子丰度是否发生改变。TAF8基因在RNA聚合酶II介导的转录过程中发挥关键调控作用。实验设计概述：本研究采用靶向RNA聚合酶II的抗体开展染色质免疫沉淀（Chromatin Immunoprecipitation，ChIP）实验，已提交患者与对照样本的免疫沉淀DNA产物，分别命名为L-IP与H-IP。为便于后续数据归一化处理，研究者向成纤维细胞染色质样品中掺入了1/10体积的果蝇染色质。此外，HFF样本的输入对照样品也将同步提交。