GSM8162371: Huh7_KLF5_CUTTAG; Homo sapiens; OTHER
ChIP-seq was performed as described previously (Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions. Methods.2009;48:240-8)；Total RNA extracted using TRIzol was used for library preparation with an Illumina mRNA sequencing (mRNAseq) sample preparation kit. Quality control of the RNA sequencing libraries was performed using a Bioanalyzer 2100 (Agilent, CA, USA) prior to paired-end sequencing on the HiSeq 3000 platform (Illumina, CA, USA); CUT&Tag was performed according to the manufacturer's protocol (Novoprotein, Shanghai, China). For the ChIPseq, input amounts of DNA was converted into high quality libraries for next-generation sequencing on the Illumina platform using the NEBNext Ultra II DNA Library Prep Kit. Bead based size selection was performed for final library size selection to obtain ChIP-seq libraries containing fragments of 250 bp to 500 bp.  For the RNA-seq, the sequencing library was prepared using an Illumina mRNA-seq sample preparation kit and was then subjected to paired-end sequencing on the Illumina HiSeq platform.  CUT&Tag was performed according to the manufacturer's protocol (Novoprotein, Shanghai, China). The DNA was mixed with a universal i5 and a uniquely barcoded i7 primer. The sample was placed in a Thermocycler for PCR, and post-PCR clean-up was performed by bead based size selection and cleanup.