GSM3770679: LA-SOL_25_1 [AC-13_S3]; Lasiodiplodia theobromae; RNA-Seq
The mycelium of each replicate (three replicates per condition) was ground in liquid nitrogen and total RNA was extracted using the Spectrum Plant Total RNA kit (Sigma), according to the manufacturer's instructions. Samples were incubated for 15 min with the DNase I digestion set (RNase-Free DNase Set, Qiagen). Integrity and quality analysis were done on a 2100 Bioanalyzer RNA (Agilent Technologies). Illumina mRNA sequencing libraries were made from 500 ng total RNA of each sample using the QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen, Vienna, Austria) according to manufacturer's protocol. Fourteen PCR cycles were used during the enrichment PCR. The size distribution, purity (absence of free adaptors) and quantity of the resulting libraries were measured using a High Sensitivity DNA chip (Agilent Technologies, Santa Clara, CA, US). The libraries were equimolarly pooled and sequenced in an Illumina Nextseq 500 high throughput flow cell, generating single-end 75bp reads.