GSM5008975: HeLa Wild Type AQ-seq 2; Homo sapiens; miRNA-Seq
Total RNA was isolated using TRIzol (Invitrogen). Total RNA was mixed with thirty synthetic miRNA-like spike-in oligos. The RNA mixture was size-fractionated by 15% urea-polyacrylamide gel electrophoresis to enrich miRNA species. Small RNA libraries were constructed using either TruSeq small RNA library preparation kit (Illumina) according to manufacturer's instructions or AQ-seq as follows: miRNA-enriched RNA was ligated to 3′ randomized adapter with 20% PEG 8000, then ligated to 5′ randomized adapter with 20% PEG 8000, reverse-transcribed, and amplified. The cDNA was sequenced using a HiSeq 2500 platform.