GSM2392544: WT late rep1 ChIPseq H3K9me3; Caenorhabditis elegans; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K9me3
Cell type
Cell type Class
Adult
Cell type
Young adult
NA
NA
Attributes by original data submitter
Sample
source_name
Whole animal, young adult
strain
N2
replicate
1
generation
F4
barcode
CGTAGA
antibody
Anti-Histone H3 (tri methyl K9)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
50 μl of the lysate was used to make IP input library. For each immumoprecipiation experiment, 2 μg of anti-H3K9me3 (ab8898, Abcam) or anti-H3K36me3 (300-95289, WAKO) was added to 400 μl of the lysate (containing approximately 50 μg DNA). The IP mix was rotated overnight at 4°C. 50 μl of Protein A(for H3K9me3) or protein G (for H3K36me3) Dynabeads (Life Technology) was added and rotated for another 2 hours. The beads were then washed three times with 800 μl ice-cold LiCl washing buffer (100 mM Tris-HCl, pH7.5, 500 mM LiCl, 1% NP-40 and 1% sodim deoxycholate). To elute the immunoprecipitation product and reverse crosslink, beads were incubated with 400 μl of worm lysis buffer (0.1 M Tris-HCl, pH 7.5, 100 mM NaCl, 50 mM EDTA, 1% SDS, and 80 μg of proteinase K) at 65 °C for 4 hours with continuous agitation (IP input lysate was treated similarly to reverse crosslink). DNA was extracted by phenolchloroform and dissolved in TE buffer. ChIP-seq libraries were made using NuGen Ovation Ultralow Library Systems v2 according to manufacturer’s instructions.