Deep sequencing of small RNAs is now the gold standard for small RNA profiling and discovery, biases in smallRNA-seq have been reported. We developed 4NBoost, a biased-minimized, quantitative miRNA and piRNA sequencing technique. Using 4NBoost, we sequenced 259 samples, including 20 tissues from mice, 18 tissues from monkeys, 4 tissues from arabidopsis thaliana, and 24 commonly used cell lines. All 4NBoost data have been integrated into our sncRNA database, SmRNAQuant, to enhance accessibility and usability.