Litchi Fruit RNA Isolation Protocol

Total RNA of pollinated and parthenocarpic litchi fruits was isolated using TRIzol reagent (Invitrogen) according to the manufacturer’s protocol^{78 (link)}. Initially, 100 mg of litchi fruit was homogenized in 1 mL of TRIzol reagent (Invitrogen) and was centrifuged at 12,000 × g for 10 min at 4 °C. The supernatant was transferred to a new clean tube, incubated at room temperature for 5 min, mixed with 0.2 mL of chloroform and shaken vigorously for 15 s. After an incubation for 3 min at room temperature, the sample was centrifuged at 12,000 × g for 15 min at 4 °C and the aqueous phase was transferred to a new clean tube. Then, 10 µg of RNase-free glycogen (Invitrogen) was added to the tube, followed by the addition of 0.5 mL of 100% isopropanol and incubation for 10 min. After the sample was centrifuged at 12,000 × g for 10 min at 4 °C, the resulting RNA sample was washed and stored at −80 °C.