GSM5216900: sRNA_intestine_1; Drosophila melanogaster; miRNA-Seq
Total RNA was prepared in triplicate with Trizol from both dissected intestines and remaining carcasses of 2- to 3-day old males reared on standard food. This RNA was size fractionated by preparative PAGE and the 18- to 29-nt long contingent were extracted. 3' and 5' linkers were then sequentially ligated onto these small RNAs using T4 RNA ligase 2, truncated K227Q (NEB) and T4 RNA ligase (NEB), respectively, and properly sized fragments were PAGE purified after each ligation. The cloned small RNAs were reverse transcribed, PCR amplified, and sequenced on a NextSeq500 (Illumina) to a depth of 10-20 million reads per library