GSM5379852: TTDV_fruit(45d)_S62; Cucumis melo; RNA-Seq
Approximately 0.25 g of the powdered, frozen tissues were mixed with 1 ml of Trizol reagent (ThermoFisher) and incubate for 5 minutes.  The solution was mixed with 0.5 ml of chloroform and centrifuged at 12,000g for 10 minutes after incubation for 5 minutes.  The aqueous phase was mixed with 350 µl of ethanol and loaded to an EconoSpin column (Epoch Life Science), and centrifuged at 8,000 g for 40 seconds.  The column was washed twice with 450 µl of 3M sodium acetate and with 320 µl of 70% ethanol.  RNA was eluted with 30 µl of RNase-free water.  Subsequently, RNA samples were treated with DNase and cleaned up with phenol-chloroform extraction and ethanol precipitation. cDNA libraries were synthesized using Universal Plus mRNA-seq with NuQuant Kit (NuGen) according to the manufacturer's protocol