Bioanalyser agilent rna 6000 pico kit

For RNA extraction, 100 mg of fresh plant material was weighed, immediately frozen in liquid nitrogen, and kept at -80 ºC until use. Three biological replicates of each one-and two-dimensional gametophytes were used for RNA-Seq. Gametophytes at specific stages were homogenized with a Silamat S5 shaker (Ivoclar Vivadent, Schaan, Liechtenstein) twice for 10 s and 5 s, respectively. Total RNA was isolated using the SpectrumTM Plant Total RNA kit (Sigma-Aldrich, Buchs, Switzerland). After DNA elimination with the TURBO DNA-free kit (Life Technologies, Carlsbad, CA), the quality of the RNA was checked using the Bioanalyser Agilent RNA 6000 Pico Kit (Agilent Technologies, Waldbronn, Germany).
Sequencing libraries were prepared using the TruSeq RNA Sample Prep Kit v2, which were sequenced on Illumina HiSeq 2000. The Transcriptome Shotgun Assembly project is available in the European Nucleotide Archive (ENA, http://www.ebi.ac.uk/ena) under the accession number PRJEB18522. The de novo transcriptome assembly in fasta format as well as the transcriptome annotation have been deposited in the Zenodo (www.zenodo.org) research data repository (https://doi.org/10.5281/zenodo.1040330).