GSM3381344: Glu72h3; Postia placenta; RNA-Seq
Total fungal RNA was extracted from mycelium with TRIzol (Life Technologies) and cleaned up with a Qiagen RNeasy Mini Kit (Qiagen, Inc.) prior to RNA sequencing or quantitative RT-PCR (qRT-PCR), a method validated in Zhang et al. (2016). For RNA-Seq, RNA was extracted using Qiagen RNeasy mini kit (cat74104), followed by genomic DNA removal and cleaning using Qiagen RNase-Free DNase Set kit (cat#79254) and Qiagen Mini RNeasy™ kit (cat#74104). Integrity of the RNA samples was assessed using the Alegient 2100 Bioanalyzer. RNA samples having RNA Integrity Number between 9 – 10 were used in this work.  Ribo-Zero rRNA removal Kit (cat#MRZPL1224) was used for enrichment of transcripts.  The SOLiD® Total RNA-Seq Kit (cat#4445374) was used to construct template cDNA for RNA-Seq following the protocol recommended by Applied Biosystems. Briefly, mRNA was fragmented using chemical hydrolysis followed by ligation with strand specific adapters and reverse transcript to generate cDNA.  The cDNA fragments, 150 to 250 bp in size, were isolated and  amplified through 15 amplification cycles to produce the required number of templates for the SOLiD™ EZ Bead™ system, which was used to generate the template bead library for ligation base sequencing by 5500xl SOLiD™ instrument (LifeTechnologies).
GEO
GSM3381344
GEO Accession
GSM3381344