GSM5746571: WGBS_OE_rep2; Oryza sativa Indica Group; Bisulfite-Seq
Cetyltrimethylammonium bromide method was used for genomic DNA extraction. Total RNA from rice leaves was extracted using TRIzol Reagent (Invitrogen, CA, USA) for RNA sequencing Genomic DNA (2-µg) spiked with 5-ng unmethylated Lambda DNA (Promega, Madison, WI, USA) was fragmented by sonication to generate fragments measuring 300–500 bp. These fragments were then ligated with 5-methylcytosine-modiﬁed adapters and subjected to bisulfide conversion using the ZYMO EZ DNA Methylation-Gold Kit (Zymo Research, Irvine, CA, USA). The bisulfide-converted DNA was purified, recycled, and then amplified by PCR with 10 cycles using KAPA HiFi HotStart Uracil + ReadyMix (Kapa Biosystems, Wilmington, MA, USA) and Illumina 8-bp index primers. RNA sequencing libraries were generated from the purified mRNA using the VAHTS Universal V6 RNA-seq Library Kit for MGI (Vazyme, Nanjing, China) following the manufacturer's recommendations with unique index codes.