sRNA Library Preparation and Sequencing

Starting total template CUSA-EV RNA (4 ng) was used to prepare the small RNA (sRNA) libraries using a QIASeq^TM miRNA Library Kit (Qiagen, Venlo, Netherlands) [104 (link)]. The sRNA library was prepared, quality control and NGS was performed by the Ramaciotti Centre for Genomics, University of New South Wales. NGS was performed on an Illumina^® NextSeq^TM 500 NGS System (Illumina, San Diego, CA, USA). The libraries were sequenced in a single-end 75 bp, high-output protocol to generate an output of up to 400 million reads at a Q30 > 85%.

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Hallal S., Ebrahim Khani S., Wei H., Lee M.Y., Sim H.W., Sy J., Shivalingam B., Buckland M.E, & Alexander-Kaufman K.L. (2020). Deep Sequencing of Small RNAs from Neurosurgical Extracellular Vesicles Substantiates miR-486-3p as a Circulating Biomarker that Distinguishes Glioblastoma from Lower-Grade Astrocytoma Patients. International Journal of Molecular Sciences, 21(14), 4954.

Publication 2020

QIASeqTM miRNA Library Kit

Library Mirna

Corresponding Organization : Royal Prince Alfred Hospital

Other organizations : National Health and Medical Research Council, The Kinghorn Cancer Centre, St Vincent's Hospital

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Variable analysis

independent variables

Starting total template CUSA-EV RNA (4 ng)

dependent variables

Preparation of the small RNA (sRNA) libraries

control variables

QIASeq™ miRNA Library Kit (Qiagen, Venlo, Netherlands)
Illumina® NextSeq™ 500 NGS System (Illumina, San Diego, CA, USA)
Single-end 75 bp, high-output protocol
Up to 400 million reads at a Q30 > 85%

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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