title: RNA-seq of ectomycorrhizal fungi
design: Fungi RNA was extracted using the TRIzol method following the manufacturer s protocol. Bulk cells or filamentous fungus are grounded to a powder using liquid nitrogen, and the powder was transferred into the 2ml tube contains 1.5ml Trizol reagent. The mix was shaked for 3min, and kept  for 5min at room temperature, then was Centrifuged at 10,000 g for 5min at 4 C. The supernatant was added 200L of chloroform/isoamyl alcohol (24:1) with 1ml lysis reagents. After centrifuged at 10,000 g for 10min at 4 C, the supernatant was transferred into another new tube with equal volume of isopropanol and put in the refrigerator at -20 C for 1h. After centrifuged at 13600 g for 20min at 4 C, the supernatant was precipitated by 1 mL of 75% ethanol and let dry for 3-5min. The RNA pellet was dissolved with 30-100L of DEPC water or RNase-free water. The concentration of the extracted RNA samples was determined using a Nanodrop system (NanoDrop, Madison, USA), and the integrity of the RNA was examined by the RNA integrity number (RIN) using an Agilent 2100 bioanalyzer (Agilent, Santa Clara, USA)