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KLF4 ChIP-seq at reprogramming 48 hours and Hic2, KLF4 ChIP-seq at reprogramming 48 hours with Hic2 overexpression

ABSTRACT: KLF4 ChIP-seq at reprogramming 48 hours and Hic2, KLF4 ChIP-seq at reprogramming 48 hours with Hic2 overexpression

PROVIDER: PRJEB64539 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR11757982_1.fastq.gz	Fastqsanger.gz
	ERR11757982_2.fastq.gz	Fastqsanger.gz
	ERR11757983_1.fastq.gz	Fastqsanger.gz
	ERR11757983_2.fastq.gz	Fastqsanger.gz
	ERR11757984_1.fastq.gz	Fastqsanger.gz

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KLF4 ChIP-seq at reprogramming 48 hours and Hic2, KLF4 ChIP-seq at reprogramming 48 hours with Hic2 overexpression

Project description:Mouse embryonic fibroblasts (MEFs) with doxycycline (Dox)-inducible reprogramming cassette MKOS-ires-mOrange and a Nanog-GFP reporter were transduced with lentiviral Dox-inducible Ty1-BFP (blue fluoresce protein with Ty1 tag in the N-terminus) or Hic2-Ty1 (Hic2 with Ty1 tag in the C-terminus) expression vector with MOI 5. One day later reprogramming was initiated by the administration of Dox. 48 hours later, the cells for Hic2 ChIP-seq with anti-Ty1 antibody were crosslinked with 2mM of Disuccinimidyl Glutarate (DSG) for 45 min at room temperature (RT) under constant agitation, before being cross-linked with 1% formaldehyde solution for 10 min at room temperature. For KLF4 ChIP-seq, cells were only cross-linked with 1% formaldehyde solution for 10 min at room temperature. ChIP-seq library was generated with the NEBNext Ultra-II DNA Library Prep kit. Each library was uniquely barcoded using NEBNext Multiplex Oligos for Illumina. Samples were sequenced with the NextSeq High 40PE.

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