RNA-Seq of Paspalum notatum sexual genotype C4-4X: floral samples at meiotic stage.
A detailed comparative transcriptomic analysis was carried out in flowers of sexual and apomictic P. notatum genotypes at different developmental stages (premeiosis, meiosis, postmeiosis and anthesis). Triplicate samples were extracted for each genotype at each developmental stage (total number of samples: 24). TruSeq Stranded mRNA Illumina libraries were generated to allow unequivocal identification of the + and - strand for each sequence (October 2017 Protocol). 1 L of each library was run in a 2100 Bioanalyzer (Agilent Technologies) using the DNA 1000 kit. Electropherograms showed that the distribution of fragments sizes were within the standard values (average 260 bp). Each library was quantified using qPCR (Light Cycler 480 Roche) and normalized to a 3 nM concentration. A single equimolar pool was prepared and quantified again using the Qiagen Library Quantification Kit. The concentration of the pooled library was 3.78 nM. The pool was used for the generation of clusters in the sequencing cell. A run generating pair ends (PE) reads of 2 x 100 bp was carried out using HiSeq-Illumina.