miRNA-Seq of isolate LGP3 : TGFb1
The library preparation was done using QIAseq miRNA preparation kit (Qiagen inc) in accordance with GCTU SOPs. In brief adapters are ligated sequentially to the 3 and 5 ends of miRNAs from 500 ng total RNA. cDNA synthesis is performed. The resulting cDNA is purified and amplified. The libraries size distribution was validated, and quality inspected on the AATI Fragment Analyser (Agilent Technologies). High quality libraries were quantified using qPCR, the concentration normalised, and the samples pooled according to the project specification (number of reads). The library pool(s) were re-quantified with qubit and optimal concentration of the library pool used to generate the clusters on the surface of a flowcell before sequencing on instrument using Nextseq500, High Output sequencing kit (76 cycles according to the manufacturer instructions (Illumina Inc.).