title: ChIP-seq of Fusarium graminearum
design: Briefly, XXX g samples were washed twice in cold PBS buffer and cross-linked with 1% formaldehyde for 10 minutes at room temperature and then quenched by addition of glycine (125 mmol/L final concentration) .Afterwards, samples were lysed with 50mM Tris-HCl (pH 8.0), 10mM EDTA, 1%SDS, 1protease inhibitor cocktail, and chromatins were obtained on ice. Chromatins were sonicated to get soluble sheared chromatin (average DNA length of 200500 bp). 20ul chromatin was saved at 20C for input DNA, and 100ul chromatin was used for immunoprecipitation by Anti-H3K4me3 antibodies (ab8580Abcam). 10 g of antibody was used in the immunoprecipitation reactions at 4 C overnight. The next day, 30 L of protein beads was added and the samples were further incubated for 3 h. The beads were next washed once with 20 mM Tris/HCL (pH 8.1), 50 mM NaCl, 2 mM EDTA, 1% Triton X-100, 0.1% SDS; twice with 10 mM Tris/HCL (pH 8.1), 250 mM LiCl, 1 mMEDTA, 1% NP-40, 1% deoxycholic acid; and twice with TE buffer 1 (10 mM Tris-Cl at pH 7.5. 1 mM EDTA). Bound material was then eluted from the beads in 300 L of elution buffer (100 mM NaHCO3, 1% SDS), treated first with RNase A (final concentration 8 g/mL) during 6 h at 65C and then with proteinase K (final concentration 345 g/mL) overnight at 45C. Immunoprecipitated DNA was used to construct sequencing libraries following the protocol provided by the NEXTFLEX ChIP-Seq Library Prep Kit for Illumina SequencingNOVA-5143-02Bioo Scientific and sequenced on Illumina Novaseq 6000 with PE 150 method.
library_name: IN_J1_R1