Small RNA-seq of Rhipicephalus (Boophilus) microplus: adult females ovaries
Total RNA or enriched small RNA fractions were prepared from eggs, larvae (unfed and frustrated), adult male and adult female as well as from adult female gut, salivary glands, and ovaries. All these samples were ground in liquid nitrogen using a sterile mortar and pestle, and then the RNA was isolated using the mirVana microRNA isolation kit according to the manufacture's instructions (Ambion, Applied Biosystems). In brief, the sequencing was performed as follows: RNA was purified by polyacrylamide gel electrophoresis (PAGE), to enrich for molecules in the range of 18--30 nucleotides, and ligated with proprietary adapters to the 5' and 3'-end termini of the RNA. The samples were used as templates for cDNA synthesis. The cDNA was amplified with 18 PCR cycles to produce sequencing libraries that were subjected to Illumina/Solexa's proprietary sequencing-by-synthesis method.
HWI-EAS157_30G31AAXX:3:1