Figure 2

Summary bench protocol for HTP96 RNA extraction. High yields of good quality RNA are isolated from as little as 100 mg of fresh tissue using a streamlined P:C-L RNA extraction method in 96-well plate format. A. Harvested tissue is frozen immediately with liquid nitrogen and homogenised using a commercial bead mill. B. Add 300 μL RE buffer, apply collection tube caps and mix briefly. C. Briefly centrifuge to collect contents, add 300 μL of P:C (pH 4.3). Apply fresh collection tube caps and mix thoroughly. D. Nucleic acids in the supernatant are precipitated with 240 μL of isopropanol and 30 μL of 3 M sodium acetate (pH 5.2) at -80°C. E. Tape the collection tubes to the rack and discard the supernatant by inverting the collection tube rack. F. Wash the nucleic acid pellet twice with 70% ethanol. G. Remove all traces of ethanol and allow the nucleic acid pellet to dry before resuspending in nuclease-free water.