Pro-Seq of MEL624 control and PCIF1 KO cells

We analyzed human melanoma cells by PRO-Seq in order to determine the effect of PCIF1 KO in Pol II dynamics PRO-seq libraries from 3 independent biological replicates were generated for Control MEL624 cells, PCIF1-KO clone 1 and PCIF1-KO clone 2, using 1 million cells per sample. Samples were spiked with 40,000 Drosophila S2 cells as a normalization spike-in control. Samples were sequenced on the NextSeq using a High Output 75-cycle kit, generating paired end 42 nt reads. Libraries were sequenced to an average read depth of 50 million mappable reads per sample.

我们通过精准转录延伸测序（PRO-Seq）分析人类黑色素瘤细胞，旨在探究PCIF1敲除（PCIF1 KO）对RNA聚合酶II（Pol II）动力学的影响。针对对照组MEL624细胞、PCIF1敲除克隆1号及PCIF1敲除克隆2号，我们分别构建了3个独立生物学重复的PRO-Seq文库，每个样本使用100万个细胞。向每个样本中掺入40,000个果蝇S2细胞，作为标准化掺入对照。采用搭载高输出型75循环测序试剂盒的NextSeq测序平台对样本进行测序，生成双端42 nt读段。所有样本的平均可比对读段测序深度达50百万条。