To annotate microRNAs (miRNAs) present in the Streptochaeta genome, we (i) sequenced sRNAs from leaf, anther and pistil tissues, (ii) compared miRNAs present in anthers to those of three other representative monocots (rice, maize, and asparagus), and (iii) validated gene targets of these miRNAs.
We collected tissues from leaf and pistil as well as 1.5, 3, and 4 mm anthers. Samples were immediately frozen in liquid nitrogen and kept at –80°C prior to RNA isolation. Total RNA was isolated using the PureLink Plant RNA Reagent (Thermo Fisher Scientific, Waltham, MA, United States). sRNA libraries were published previously (Patel et al., 2021 (link)). RNA sequencing libraries were prepared from the same material using the Illumina TruSeq stranded RNA-seq preparation kit (Illumina Inc., United States) following manufacturer’s instructions. Parallel analysis of RNA ends (PARE) libraries were prepared from a total of 20 μg of total RNA following the method described by Zhai et al. (2014) (link). For all types of libraries, single-end sequencing was performed on an Illumina HiSeq 2000 instrument (Illumina Inc., United States) at the University of Delaware DNA Sequencing and Genotyping Center.
We collected tissues from leaf and pistil as well as 1.5, 3, and 4 mm anthers. Samples were immediately frozen in liquid nitrogen and kept at –80°C prior to RNA isolation. Total RNA was isolated using the PureLink Plant RNA Reagent (Thermo Fisher Scientific, Waltham, MA, United States). sRNA libraries were published previously (Patel et al., 2021 (link)). RNA sequencing libraries were prepared from the same material using the Illumina TruSeq stranded RNA-seq preparation kit (Illumina Inc., United States) following manufacturer’s instructions. Parallel analysis of RNA ends (PARE) libraries were prepared from a total of 20 μg of total RNA following the method described by Zhai et al. (2014) (link). For all types of libraries, single-end sequencing was performed on an Illumina HiSeq 2000 instrument (Illumina Inc., United States) at the University of Delaware DNA Sequencing and Genotyping Center.