Centre : Central Tuber Crops Research Institute(CTCRI),TVM
Name of the Protocol
Total RNA isolation from taro/P. colocasiae
Nature : Modification of existing protocol
Crop / Pathogen : taro/P. colocasiae
The harvested fungal mycelia/ plant material was ground to a fine powder in liquid N2, then 2ml TRIzol Reagent (Invitrogen) was added and the sample ground further until the slurry had thawed. The sample was split into two equal volumes and transferred to 2ml Eppendorf tubes. After 5 min incubation at room temperature (RT) 300 μl of chloroform was added, the tube shaken vigorously for 15 seconds and allowed to stand at RT for 3 minutes. The sample was then centrifuged at 13,000 × g for 15 minutes at 4°C. The supernatant from the final extraction step was transferred to a clean 1.5 ml Eppendorf tube and the RNA precipitated with 500 μl isopropanol at −20°C for 2 hours or longer (over night when small RNAs are to be recovered). Precipitated RNA was collected by centrifugation at 13,000 × g for 15 minutes at 4°C, the pellet washed with 1ml of ice cold 75% ethanol and air dried briefly at RT. The RNA pellets were resuspended in 20 μl of nuclease free water and the two duplicate tubes combined.