Pou5f3 and Sox2 ChIP-seq
Zebrafish homologues of the mammalian core pluripotency factors, Pou5f3 and SoxB1 are also crucially involved in Zygotic Genome Activation in zebrafish. To identify the components of Pou5f3 and SoxB1-dependent transcriptional networks, Wolfgang Driever lab determined the genomic binding sites for Pou5f3 at 512 cell stage and 50% epiboly and Sox2 at 50% epiboly stage using ChIP-seq. For Pou5f3 Chip-seq, MZspg mutant embryos, injected with 25 pg/embryo synthetic Flag-Pou5f3 mRNA and anti-Flag antibody were used. For Sox2 ChIp-seq, AB/TL zebrafish embryos and anti-Sox2 antibody were used. 76-basepair single-end reads of 50% epiboly Pou5f3 (twice, to reach the deeper read density) and Sox2 ChIP-seq and input control were sequenced the Genome Analyzer IIx (Illumina), resulting in 4 FASTQ files for ChIP-seq: one for Sox2 and for Input control, and two for Pou5f3. For the pre-MBT Pou5f3 ChIP samples, 51-basepair single-end reads were sequenced by GATC Biotech AG using a HighSeq 2000 (Illumina), resulting in 1 additional FASTQ file for ChIP-seq
May 30, 2016, 5:20 a.m.
Nov. 7, 2018, 5:15 a.m.
|Sequencing ID||Technical Replicate||Sequencing Lab||Platform||Instrument||Strand||Sequencing Date|
|DCD000973SQ||DCD001183TR||Driever Lab, University of Freiburg||Illumina||GAIIx||unstranded||05/05/2012|