title: GSM1468703: (III) polII_ChIP__N2A__Luc_KD; Mus musculus; ChIP-Seq
library_construction_protocol: Lysates were clarified after micrococcal nuclease treatment and RNA-Poymerase II-DNA complexes were isolated with antibody. Libraries were prepared according to Life Techonology's instructions accompanying the Ion XpressTM Plus gDNA Fragment Library Preparation Kit. Briefly, DNA fragment was treated with the mixture of T4 DNA Polymerase, DNA Polymerase I, Large (Klenow) Fragment, T4 Polynucleotide Kinase using T4 DNA Ligase Buffer (New England Biolabs, Ipswich, MA, UK). ChIP-seq Libraries were prepared from the pretreated ChIP DNA using the Life Techonology Ion XpressTM Plus gDNA Fragment Library Preparation Kit. Ligation products were size-selected by LabChip XT and LabChip XT DNA 750 Assay Kit (PerkinElmer, Waltham, MA, USA). After purifying constructed libraries with Agencourt AMPure XP (Beckman Coulter, Brea, CA, USA), high-throughput sequencing was performed using the Ion Proton System.