GSM1191102: Nymph; Panonychus citri; RNA-Seq
More than 3000 embryos, 2500 larvae, 1500 nymphs (a mixture of protonymphs and deutonymphs) and 1000 adults were gathered into four centrifuge tubes, respectively, and stored at -80 °C for RNA extraction. Total RNA from the four samples was extracted using the TriZol reagent (TransGen Biotech, Beijing, China), according to the manufacturer's instructions. For each library, small RNAs of 18 to 30 bp in length were separated by 15% polyacrylamide gel electrophoresis (PAGE). The ligation between these small RNAs and 5' and 3' adaptors was performed using T4 RNA Ligase (Takara:D2050). The 5'- and 3'-ligated RNA products were purified and then reverse transcribed for the first-strand cDNA using a SuperScript® II Reverse Transcriptase kit (InvitrogenTM: 18064-014) with the complementary sequence of the 3' adaptor. Thereafter, the cDNAs were used as templates for double-stranded cDNA synthesis using PCR amplification in a Veriti Thermal Cycler (Applied Biosystems), using primers that anneal to the adapters. After second-strand cDNA synthesis, the adaptor sequence primers were used to amplify the PCR products as a small RNA library for deep sequencing.
GEO Accession
GSM1191102