Verso cdna synthesis transscript kit

E.Z.N.A.® Tissue DNA Kit (Omega Bio-tek., Norcross, GA, United States) was used to extract DNA from insect individuals. Total plant RNA was extracted from citrus leaves using E.Z.N.A.® Plant RNA Kit (Omega Bio-tek., Norcross, GA, United States). Total insect RNA was extracted from a single D. citri using TRlzol® Reagent (Life Technologies, Guangzhou, China). The concentration and purity of total DNA or RNA samples were quantified by absorbance using NanoDrop™ One micro-volume UV–Vis spectrophotometer (Thermo Scientific, Shanghai, China). The total RNA samples were individually used for reverse transcription using Verso cDNA Synthesis (TransScript) Kit (TransGen Biotech, Beijing, China). cDNA samples were stored at −80°C for further use.

The RNA from each D. citri was extracted individually using TRlzol^® Reagent (Life Technologies, Guangzhou, China). RNA from the midribs of either citrus leaves or orange jasmine leaves was isolated using the TransZol Plant kit (TransGen Biotech, Beijing, China). Genomic DNA contamination was eliminated by digestion with RNase-free DNase I (TaKaRa, Shuzo, Kyoto, Japan). Total RNA concentration was estimated with fluorescence-based Qubit™ (Shanghai, China). The purity of total RNA was determined by absorbance using NanoDrop™ One (Thermo Scientific, Shanghai, China), where RNA samples with OD260/OD230 2.0–2.4 and OD260/OD280 of 1.8−2.2 were further reverse transcripted to cDNA. cDNA was synthesized from 500 ng of total RNA using Verso cDNA Synthesis (TransScript) kit (TransGen Biotech, Beijing, China) with random hexamers primers following the manufacturer’s instructions. The obtained cDNA samples were used as templates for PCR or qPCR amplification, as described below.

Total plant RNA was extracted from the midrib of either citrus leaves or orange jasmine leaves using plant tissue E.Z.N.A.^® HP plant RNA extraction kit (Omega Bio–tek., Norcross, GA, USA). Genomic DNA contamination was eliminated by digestion with RNase-free DNase I (TaKaRa Bio–tek., Shuzo, Kyoto, Japan). Total insect RNA was extracted from tissues of a single D. citri using TRlzol^® Reagent (Life Technologies, Guangzhou, China). The concentration and purity of total RNA were determined by absorbance using NanoDrop™ One (Thermo Scientific, Shanghai, China). RNA samples with OD260/OD230 of 2.0–2.4 and OD260/OD280 of 1.8−2.2 were selected and stored at −80 °C for further use. The total RNA samples were individually used for reverse transcription by Verso cDNA Synthesis (TransScript) kit (TransGen Biotech, Beijing, China).