MiRNA-Seq of human sample: PBMCs of Type 1 Diabetic without complications
The production of small RNA libraries was performed using 1g/5L of total PBMCs RNA with the aid of the TruSeq Small RNA Library Preparation Kit (Illumina, San Diego, CA). The samples were pooled (8 per pool) and the 147-nucleotide miRNA bands were size-selected using 7% polyacrylamide gel electrophoresis (PAGE), running for 90 minutes at 145V. Subsequent to gel purification, using a 5 m filter tube (IST Engineering, Milpitas, CA), the pooled samples were quantified by RT-qPCR, using the KAPA Library Quantification Kit (KAPA Biosystems, Wilmington, MA) and sequenced using MiSeq Reagent Kit v2 (50 cycles) on the MiSeq system (Illumina).