miRNA Library Preparation and Sequencing

Preparation of miRNA libraries and sequencing was performed as described by Koppers-Lalic et al. [28 (link),29 (link)]. After RNA isolation, samples were adjusted to have the same amount of total RNA concentration (500 pg in 7 μL). Libraries were prepared using the standard small-RNA Library Prep Kit for Illumina. To assess samples’ quality before sequencing, the concentration of the samples was determined using the Fragment Analyzer and all samples met the quality requirements. Sequencing was performed using the HiSeq 4000 instrument (Illumina, San Diego, CA, USA) with 150 bp paired-ends.

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Mantini G., Meijer L.L., Glogovitis I., In ‘t Veld S.G., Paleckyte R., Capula M., Le Large T.Y., Morelli L., Pham T.V., Piersma S.R., Frampton A.E., Jimenez C.R., Kazemier G., Koppers-Lalic D., Wurdinger T, & Giovannetti E. (2020). Omics Analysis of Educated Platelets in Cancer and Benign Disease of the Pancreas. Cancers, 13(1), 66.

Publication 2020

small-RNA Library Prep Kit HiSeq 4000

Isolation Library Mirna

Corresponding Organization : Fondazione Pisa

Other organizations : Plovdiv University, Scuola Superiore Sant'Anna, University of Pisa, University of Surrey

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Variable analysis

independent variables

Preparation of miRNA libraries and sequencing protocol

dependent variables

MiRNA expression levels

control variables

Total RNA concentration (500 pg in 7 μL) of samples
Quality requirements for samples before sequencing

controls

Positive control: None mentioned
Negative control: None mentioned

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