GSM2601577: BCGiv_NI_input_rep2; Mus musculus; ChIP-Seq
DNA was extracted using the PureGene DNA extraction kit (Gentra Systems). Samples from BCG­vaccinated and non-vaccinated, Mtb-infected and non-­infected BMDMs were crosslinked with 37% w/v formaldehyde for 20 minutes at RT and immediately quenched for 5 minutes with 1M Glycine at RT. The formaldehyde fixed samples were then sonicated to 150­500 bp using an E220 sonicator (Covaris), and subsequently ChIP-­DNA was prepared using a manual chromatin immunoprecipitation method with Antibodies­Antigen incubation for 18 hours, followed by bead incubation for 150 minutes, and 7 5­minute washing steps. Approximately 1x106 cells were used for each ChIP and ~50,000 cells for the input. The following antibodies were used: H3K4me1 (Company: CST, Cat. No.: 5326P, Lot No.: 1), H3K4me3 (Thermo Fisher Scientific, MA5-­11199), H3K27ac (Abcam, Ab4729, Lot No.: GR200563­1). ChIP and Input libraries were prepared using the MicroPlex Library Preparation Kit v2 (C05010013, Diagenode), with alterations including: PCR enrichment (9 to 18 cycles) prior to size selection, and use of Ampure beads for size selection (250-­350 bp).
GEO Accession
GSM2601577