title: GSM6250439: RNA_sDH_rep1; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: Total RNA from 6th, 7th and 8th leaves was extracted with a RNeasy Plant Mini kit (Qiagen). For RNA-seq analysis, the total RNA was treated with a Poly(A) mRNA Magnetic Isolation Module Kit (NEB #E7490). DNA of 6th, 7th and 8th leaves was purified using a DNeasy® Plant Mini Kit (Qiagen) and eluted in 150 µL of elution buffer. Experiments were performed for two biological replicates. RNA-seq Libraries were prepared from the resulting mRNA with a NEBNext Ultra Kit (NEB #E7530). The libraries were constructed according to a standard protocol (Illumina). The RNA-seq libraries were sequenced at Novogene via a Novaseq instrument in 150-bp paired-end mode. For BS-seq libraries, The DNA was fragmented into 200-300 bp fragments using Covaris S220 and later Bioruptor® Plus from Diagenode (60 cycles, 30s on, 30s off). After adaptor ligation, bisulfite treatment was performed using the NEBNext® Enzymatic Methyl-seq Kit. PCR amplification of the library molecules was performed using Q5U Master Mix DNA polymerase (NEBNext®). Libraries were sequenced at Novogene (Hong kong, China) on a NovaSeq instrument in 150-bp paired-end mode. Experiments were performed on two biological replicates.