GSM3450345: HepG2 HA-METTL14 rep2 (ChIP-seq); Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
Epitope tags
Cell type
Cell type Class
Liver
Cell type
Hep G2
Primary Tissue
Liver
Tissue Diagnosis
Carcinoma Hepatocellular
Attributes by original data submitter
Sample
source_name
HepG2 cells
cell line
HepG2
ip antibody
HA antibody (11867423001, Roche)
replication
2
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
METTL14 ChIP experiment was performed using iDeal ChIP-seq Kit for Transcription Factors (Diagenode) following the manufacturer's protocol. Briefly, HepG2 cells transduced with HA-tagged METTL14 were crosslinked with formaldehyde, lysed, and subjected to chromatin shearing by sonication in a Bioruptor Pico (Diagenode) with 30s on/30s off for 20 cycles. Immunoprecipitation was performed using 2 µg anti-HA antibody (11867423001, Roche). Product DNA (including IP product and the Input control) was decrosslinked and subjected to library construction using KAPA Hyper Prep Kit (Roche). Libraries were purified using AxyPrep Mag PCR Clean-up kit (Thermo Fisher Scientific). Each library was quantified using a Qubit fluorometer (Thermo Fisher Scientific) and the size distribution assessed using the 2100 Bioanalyzer (Agilent Technologies). The amplified libraries were hybridized to the Illumina single end flow cell and amplified using the cBot (Illumina). Single end reads of 50 nt were generated on Illumina HiSeq2500.