Qubit rna assay kit in qubit 3

WT and oslil3 mutant plant samples were collected at the L3 seedling stage and whole leaf samples were harvested for RNA extraction (3 biological replicates with 6–8 individual plants for each replicate). Leaves from oslil3 and WT were rapidly frozen in liquid nitrogen and total RNA was extracted with the EZNA plant RNA kit (Omega Bio-Tek, GA, USA) according to the manufacturer’s instructions. RNA quantity and purity were checked by the kaiaoK5500 Spectrophotometer (Kaiao, Beijing, China). RNA integrity and concentration were analyzed with the RNA Nano 6000 Assay Kit of Bioanalyzer 2100 (Agilent Technologies, CA, USA). RNA concentration for library construction was measured using Qubit RNA Assay Kit in Qubit 3.0 (Life Technologies, Grand Island, NY, USA). Dilutions were prepared at a concentration of 1 μg·μl^− 1.

RNA concentration of library was measured using Qubit RNA Assay Kit in Qubit 3.0 (Life Technologies) for quantification and then diluted to 1 ng/μL. Insert size was assessed using the Agilent Bioanalyzer 2100 system. After the insert size met the requirement, the CFX 96 fluorescence quantitative PCR instrument (Bio-Rad, CA, USA) was used to quantify the library effective concentration (library effective concentration >10 nm) using Bio-Rad iQ SYBR GRN Kit.