Transcriptome Profiling of H. monotropa

The procedures for sample collection, DNA and RNA library preparation for most datasets (DNA shotgun and mate-pair libraries and an inflorescence transcriptome library) and sequencing were described in the reports of Logacheva et al. (2016) (link) and Schelkunov, Penin & Logacheva (2018) (link). In addition to the data generated in previous studies, we used four new datasets that represent the transcriptomes of the anthers and ovules of H. monotropa. The samples were collected in the same location as previous samples but in 2018. H. monotropa is not an endangered or threatened plant species; thus, no specific permissions were required for its collection. RNA was extracted using an RNEasy kit (Qiagen, Hilden, Germany) with the addition of the Plant RNA Isolation Aid reagent (Thermo Fisher, Waltham, MA, USA) to the lysis buffer. The removal of ribosomal RNA was performed using a Ribo-Zero plant leaf kit (Illumina, San Diego, CA, USA), and further sample preparation was performed using a NEBNext RNA library preparation kit (New England Biolabs, Ipswich, MA, USA). The libraries were sequenced on the HiSeq 4000 platform (Illumina, San Diego, CA, USA) in 150-nt paired-end mode at the Skoltech Genomics Core Facility. The reads were deposited in the NCBI Sequence Read Archive under BioProject PRJNA573526.