GSM7847122: Mycorrhizal, 5 weeks-post-inoculation, biol rep 1; Marchantia paleacea; RNA-Seq
Samples were harvested after 5, 8 and 11 WPI, washed in distilled water and fixed in ice-cold 100% methanol. For each biological replicate, four thalli were collected and pooled together for subsequent analysis. The midribs of collected thalli were excised to remove apical notches, lateral margins and gemma cups. After sectioning, midribs were washed 3x in ice cold phosphate-buffered saline (pH 7.4) and snap frozen in liquid nitrogen. The excised midribs were ground in liquid nitrogen with mortar and pestle and total RNA was extracted using PureLinkTM Plant RNA Reagent (Thermo Fisher Scientific, Waltham, USA), following manufacturer's instructions. 2% (w/v) polyvinylpyrrolidone (PVP40) was added to the Plant RNA Reagent mixture before extraction to improve RNA purity. cDNA library preparation was performed using 1.1μg of total RNA with a TruSeq Stranded mRNA Library High Throughput Kit (Illumina, San Diego, USA) according to manufacturer's instructions (Catalog #RS-122-9004, DOC, Part #15031047 Rev.E). Library quality was assessed using a DNA1000 chip on a Bioanalyzer 1200 (Agilent Technologies, Santa Clara, USA) and library quantities were measured with a Qubit dsDNA BR Assay Kit (Thermo Fisher Scientific, Waltham, USA).