title: GSM2934977: tRNA-iMetCAU in NSun2 Null; Drosophila melanogaster; Bisulfite-Seq
library_construction_protocol: Flies (10 females and 10 males) were mechanically homogenized in TriReagent (Sigma) using a pestle and total RNA was isolated. Bisulfite conversion was performed on 1 µg RNA after DNaseI treatment using the EZ RNA Methylation™ Kit (ZymoSearch). Bisulfite-treated RNA was reverse transcribed with bisulfite-specific and tRNA-specific primers incorporating an internal 4-letter barcode preceded and succeeded by two randomized nucleotides (NNXXXXNN) to follow PCR biases using the SuperScript III First-Strand Synthesis System (Thermo Fisher Scientific). Amplicons for Illumina sequencing were generated by two sequential PCR and sequencing was performed on an Illumina miSeq platform.
attributes.tag: GEO Accession
attributes.value: GSM2934977