Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Epitope tags

Cell type

Cell type Class
Larvae
Cell type
L4
NA
NA

Attributes by original data submitter

Sample

source_name
whole worm
strain background
N2
genotype/variation
LONP1-FLAG
developmental stage
L4 stage hermaphrodite worms
chip antibody
Monoclonal ANTI-FLAG M2 antibody (Sigma, F1804)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Synchronized worms were cultured in liquid and harvested at early L4 stage by sucrose flotation. The worms were lysed via Teflon homogenizer in cold PBS with protease inhibitors (Roche). Cross-linking of DNA and protein was performed by treating the worms with 1.85% formaldehyde for 15 min. Glycine was added to a final concentration of 125mM for 5 min at room temperature to quench the formaldehyde. The pellets were resuspended twice in cold PBS with protease inhibitor. Samples were sonicated in Bioruptor (Diagenode) for 15 min at 4°C on high intensity (30s on and 30s off). Samples were transferred to microfuge tubes and spun at 15,000*g for 15 min at 4°C. The supernatant was precleaned with pre-blocked ChIP-grade Pierce™ magnetic protein A/G (Thermo Scientific) and then incubated with Monoclonal ANTI-FLAG® M2 antibody (Sigma, F1804) or Mouse mAb IgG1 Isotype Control (Cell Signaling Technology, G3A1) rotating overnight at 4°C. Then, the antibody-chromatin complex was precipitated with magnetic beads or protein A sepharose beads (Invitrogen) dependent on the host of generated antibodies. After washing, the crosslinks were reversed by incubation at 65 °C overnight and further treated with RNaseA 37 °C for 1 hour and then proteinase K 55 °C for 2 hours, respectively. Finally, immunoprecipitated and input DNA were purified with ChIP DNA Clean & Concentrator (Zymo Research, D5205) and used as templates for qPCR or next generation sequencing.

Sequencing Platform

instrument_model
Illumina MiSeq

ce11

Number of total reads
26256039
Reads aligned (%)
62.2
Duplicates removed (%)
7.6
Number of peaks
363 (qval < 1E-05)

ce10

Number of total reads
26256039
Reads aligned (%)
62.2
Duplicates removed (%)
7.6
Number of peaks
363 (qval < 1E-05)

Base call quality data from DBCLS SRA