PRJNA855900
Transcriptome sequencing of pepper anthers
The wild-type pepper cultivar has normal pollen fertility and purple anthers. A mutant with lower pollen fertility, rpf1, was obtained from its progeny by EMS mutagenesis,Total RNA was extracted using the TRIzol method from three replicate anther samples of wild-type and mutant rpf1 plants just before opening, and total RNA integrity was measured on an Agilent 2100 Bioanalyzer using an RNA Nano 6000 Assay kit. Oligo(dT) magnetic beads were used to enrich mRNA, which was then fragmented using fragmentation buffer. After reverse transcription into cDNA, AMPure XP beads were used for purification and repair, and appropriate size fragments were selected for PCR enrichment to construct a cDNA library for sequencing on the Illumina HiSeq 4000 platform.