Rna nano 6000 assay kit of the agilent bioanalyzer 2100 system

The total RNA of bermudagrass leaf was extracted by Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. The NanoPhotometer^® spectrophotometer (IMPLEN, Westlake Village, CA, USA) was applied to check the RNA purity. To guarantee the RNA purity, degradation, and contamination of RNA were monitored on 1% agarose gels. The Qubit^® RNA Assay Kit was applied to measure the RNA concentration in Qubit^® 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA). The RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, Santa Clara, CA, USA) was applied to detect the RNA integrity.

Tissues were ground thoroughly using a grinder after being melted at room temperature, followed by the extraction of total RNAs by TRIzol reagent (Takara, China), following the protocol of the manufacturer. RNAs were not degraded and contaminated, which was confirmed by 1% agarose gels. The NanoPhotometerVR spectrophotometer [IMPLEN, the United States of America (United States)] was utilized for testing the purity of RNAs. Then, a QubitVR RNA Assay Kit in QubitVR 2.0 Flurometer (Life Technologies, United States) was used for measuring the concentration of RNAs. The RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2,100 system (Agilent Technologies, United States) was applied to assess the integrity of RNAs within a RIN value ranging between 6.0 and 8.0. The above RNAs were utilized for the sequencing of small RNAs and quantitative real-time polymerase chain reaction (qRT-PCR) analysis.