ABSTRACT
Premise of the study: We describe a field and lab workflow developed for plant phylotranscriptomic projects, involving field collected cryogenic tissues, RNA extraction and quality control, and library preparation. We also make recommendations for sample curation.
Methods and Results: 216 frozen tissue samples of Caryophyllales and other angiosperm taxa were collected from the field or botanical gardens and were subjected to RNA extraction, stranded mRNA library preparation and sequencing on Illumina HiSeq platforms. These include difficult mucilaginous tissues such as those of Cactaceae and Droseraceae.
Conclusions: Our workflow is not only cost effective (~$270 per sample, as of August 2016, from tissue to reads) and time efficient (~5 hours a sample including all lab work and sample curation), but has proven robust for extraction of difficult samples such as tissues containing high levels of secondary compounds.
