GSM8752021: METTL3, CHIP, Control; Homo sapiens; ChIP-Seq
Total RNA was purified from cells using the TRIzol reagent (TIANGEN, cat# DP424), followed by standard chloroform phase separation and alcohol precipitation. cDNA was synthesized from 1 μg total RNA, and reverse transcription was performed using the HiScript II Q RT SuperMix kit (Vazyme, cat#R222-01) according to the manufacturer's instructions. The ATAC-seq assay was performed as described previously. Brieﬂy, 5x104 cells were washed twice with PBS and lysed in lysis buffer containing 10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, and 0.1 % NP-40. The lysate was washed in PBS and treated with Tn5 transposase at 37°C for 30 minutes. DNA was puriﬁed using the DNA purification Kit (TIANGEN, Cat# DP214). DNA library was then generated by Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme, cat# TD711-01). RNA sequencing libraries were prepared from 1 μg of total RNA using the VAHTS® Universal V8 RNA-seq Library Prep Kit for Illumina (Vazyme, cat# NR605-01/02) following the manufacturer instructions.  ChIP-DNA libraries were prepared using the Illumina's VAHTS Universal Pro DNA Library Prep Kit according to the manufacturer's instructions (Vazyme, cat# ND608-01/02).  The ATAC-seq DNA was puriﬁed using the DNA purification Kit (TIANGEN, Cat# DP214). ATAC chromatin DNA library was then generated by Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme, cat# TD711-01). Hi-C assay was performed to generate a genome-wide interaction as described previously with Arima-HiC Kit (Cat: A410030) (https://arimagenomics.com/) with minor modifications.