GSM4212840: ChIp-seq_Pfhmgb1_KO_h3k9ac_rep2; Plasmodium falciparum; ChIP-Seq
Nuclei were isolated by Lysis Buffer and Dounce homogenization, followed by sonication to generate 200–600 bp fragments in length on the highest power setting for 10-20 min at 30s intervals. Protein-DNA complexes are selectively immunoprecipitated using 3 µg of Rabbit anti-H3K9me3 antibody (Abcam, ab8898), 3 μg of Rabbit anti-H3K9Ac antibody (Millipore, 07-352), or rabbit IgG (Sigma) as control. Formaldehyde induced cross-links are reversed with NaCl and DNA extracted using Proteinase K and MinElute PCR purification kit (Qiagen, 28006). The ChIP DNAs were used to prepare sequencing libraries, including end-repair (Invitrogen No.16123), adding protruding 3' A base (NEB No.M0212L), adapter ligation(NEB No.M2200L), size selection with Ampure beads (for 250 bp inserts) and amplification of libraries(KAPA Biosystems KB2500) (PCR program: 1 min at 98 ℃ followed by 12 cycles of 10 s at 98 ℃, 1 min at 65 ℃; finally extended 5 min at 65 ℃). Amplified libraries were cleand with Ampure beads The library were sequenced on an Illumina HiSeq Xten systerm with PE150 strategy.
GEO Accession
GSM4212840