High sense dna chips

Gel purified libraries were quantified using Agilent 2100 Bioanalyzer with High-Sense DNA chips. Equimolar amounts of libraries were pooled and quantified by Bioanalyzer with High-Sense DNA chips. Pooled libraries were normalized and denatured at a working concentration range of 6–8 pM with 5% PhiX spike-in for flow cell cluster generation. Samples were sequenced until new miRNA detection plateaued for each tissue (Supplementary Figure S1).

Small RNA libraries were generated using NEXTflex Small RNA Library Prep Kit v2 (BIOO) with 16 cycles of PCR amplification. Amplified libraries were resolved on a 6% TBE gel for size selection. The 140- to 160-nucleotide bands corresponding to adapter-ligated libraries were excised and recovered in a DNA elution buffer followed by ethanol precipitation and resuspension in 11 μL of ultra-pure water. Gel purified libraries were quantified using Agilent 2100 Bioanalyzer with High-Sense DNA chips. Equimolar amounts of libraries were pooled and quantified by Bioanalyzer with High-Sense DNA chips. Pooled libraries were normalized and denatured at a working concentration range of 6–8 pM with 5% PhiX spike-in for flow cell cluster generation.