Transcriptome analysis of Thga1 mutants of Trichoderma harzianum Th-33 in conidiation
Total RNA was extracted using TruSeq RNA Sample Preparation Kit v2-Set A following the manufacturer’s protocol. The RNA quality and quantity were determined using a Agilent 2100 BioAnalyzer. The QUBIT RNA ASSAY KIT was used for accurate quantification of the initial total RNA. After total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT) were used to isolate mRNA. The mRNA was fragmented into short fragments when it was mixed with the fragmentation buffer. Then, cDNA was synthesized using the SuperScript II reverse transcriptase following the manufacturer’s protocol. Afterwards, the short fragments were connected with adapters. The suitable fragments were selected as templates for PCR amplification. During the QC steps, a 2100 Bioanalyzer High Sensitivity DNA chip was used for quantification and qualification of the sample library. Paired-end libraries were sequenced using Illumina HiSeqTM 2000 system.