GSM2053304: Odd-Tug1-PD2; Mus musculus; OTHER
Gluteraldehyde crosslinked cells were subjected to nuclear extraction protocol and nuclei were subjected to sonication by BioRuptor. 1% Input Chromatin was set aside and Chromatin was incubated with biotynilated Tug1-Even or Tug1-Odd oligos, DNA was reverse-crosslinked and purified by ethanol precipitation. 10 ng of DNA samples were prepared for Illumina sequencing as the following steps: 1) DNA samples were blunt-ended with T4 DNA polymerase and Klenow polymerase; 2) a dA base was added to the 3' end of each strand by Klenow (exo minus) polymerase; 3) Illumina's genomic adapters were ligated to the DNA fragments; 4) PCR amplification was performed to enrich ligated fragments; 5) Enriched product of ~200-400bp was cut out from gel and purified with QIAquick Gel Extraction Kit. The completed libraries were quantified by Agilent 2100 Bioanalyzer. The libraries were denatured with 0.1 M NaOH to generate single-stranded DNA molecules, captured on Illumina flow cell, amplified in situ. The libraries were then sequenced on the Illumina HiSeq 2000 following the TruSeq Rapid SBS Kit protocol.
GEO Accession
GSM2053304