GSM4476685: 17-day embryo AQ-seq; Mus musculus; miRNA-Seq
Commercially available: Mouse Total RNA Master Panel; Takara AQ-seq (bias-minimized sRNA-seq) libraries were constructed using total RNAs from fifteen mouse tissues (Mouse Total RNA Master Panel; Takara) as described in our previous study (Kim et al., 2019). Briefly, we mixed 5 μg of total RNAs with 10 fmole of thirty equimolar spike-in RNAs which are miRNA-like non-human/mouse/frog/fish RNAs used for bias evaluation. Small RNAs were enriched by size fractionation by 15% urea-polyacrylamide gel electrophoresis and sequentially ligated to the randomized adapter at the 3′ and 5′ ends. The ligated RNAs were reverse-transcribed using SuperScript III reverse transcriptase (Invitrogen), amplified using Phusion High-Fidelity DNA Polymerase (Thermo Scientific), and subjected to high-throughput sequencing on the MiSeq platform (Illumina).