Illumina HiSeq 2000 sequencing; Collection of various small RNA sequencing libraries of C. Reinhardtii.
Collection of various small RNA sequencing libraries of C. Reinhardtii.
C. reinhardii strains were obtained from the Chlamydomonas Resource Center (University of Minnesota) and maintained by stricking cells into new fresh solid media every two months. Media 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS)-acetate-phosphate (TAP) media (Harris 2009) in the presence of 15% agar. C. reinhardtii was grown in liquid TAP for a few days with constant shaking at 25 ᴼC under continuous illumination. Cells were harvested when cultures reached saturation Total RNA was extracted with Trizol reagent by following a protocol previously described (Molnar et al., 2007, Nature). Cloning of small RNAs was performed directly from total RNAs by using the Illumina TruSeq v2 RNA Sample Preparation Kit