RNA extraction protocol (Trizol)
Add 1000 µL of Trizol per 50-100 mg of parsite tissues in a 2 mL eppendorf tube (RNAse free) and place the parasite tissues into their respective, labelled tube. 
Add 1 metal bead into each tube and place the tubes into a TissueLyzer.
Run it at 30 Hz for 3 minutes. 
Centrifuge at 12,000 x g for 10 minutes at 4°C. 
Discard top layer of liquid formed in the eppendorf after the centrifugation (contains fatty acids). 
Transfer supernatant into a new and labelled eppendorf tube (2 mL).
Incubate at room temperature for 5 minutes.
Add 200 µL of chloroform per tube. 
Mix vigorously by inverting the tubes up and down for 1 minute.
DO NOT VORTEX!
Centrifuge at 12,000 x g for 15 minutes at 4°C. 
Transfer with precaution the aqueous phase (supernatant) into a new 2 mL eppendorf tube. 
Add 500 µL of RNAse free isopropanol (100%) and mix thoroughly.  
Incubate for 15 minutes at room temperature.  
Centrifuge at 12,000 x g for 10 minutes at 4°C. 
Discard supernatant. 
Add 1000 µL of 75% ethanol (kept cold at -20°C) per tube. 
Vortex thoroughly and centrifuge at 7500g for 5 minutes at 4°C.
Discard supernatant and let the tubes dry out with the cap opened for 5-10 minutes. 
Add 20-50 µL of DEPC treated water into each tube. 
Place the tubes into the incubator (65°C) for 1 minute. 
Vortex thoroughly and repeat step 19 until RNA is completely dissolved.
