Next ultra 2 rna library prep kit

For expression profiling, total RNA was isolated from two biological replicates of 3-week-old Arabidopsis Col-0 plants using TRIzol according to the manufacturer’s protocol. The amount and quality of RNA were tested with an Agilent Bioanalyzer 2100 system and a Qubit RNA Assay Kit (Life Technologies). Ribosomal RNA was removed by using an mRNA Miniprep Kit (Sigma, MRN10). An NEB Next Ultra II RNA Library Prep Kit for Illumina (NEB, E7770S) was used to construct the library, and sequencing was performed on an Illumina NovaSeq6000 platform with 150-bp paired-end reads at Annoroad Gene Technology Co., Ltd. (Beijing, China). The clean reads were aligned to TAIR10 using TopHat v2.0.9 [51 (link)]. Cufflinks v2.1.1 was used to calculate the FPKMs of the coding genes [52 (link)]. Differentially expressed genes (DEGs) were called using Cuffdiff (v2.1.1) with an FDR < 0.05.

Total RNA of Arabidopsis and rice were isolated with TRIzol (15596026, Thermo Fisher, Waltham, MA, USA). Ribosomal RNA was removed by using an mRNA Miniprep Kit (Sigma, MRN10). An NEB Next Ultra II RNA Library Prep Kit for Illumina (NEB, E7770S) was used to construct the library, and sequencing was performed on an Illumina HiSeq 2000 platform with 100-bp paired-end reads at Jiangbei New Area Biopharmaceutical Public Service Platform Co., Ltd. (Nanjing, China).