path: root/general/datasets/Tigem_hg38_ret_rna_seq_0916/experiment-design.rtf

<h3>RNA extraction, library preparation and sequencing</h3>

<p>Total RNA was extracted from the 50 human retina samples using the miRNeasy Kit (QIAGEN) according to the manufacturer&#39;s instructions. RNA was quantified using a NanoDrop ND-8000 spectrophotometer (NanoDrop Technologies) and the integrity was evaluated using an RNA 6000 Nano chip on a Bioanalyzer (Agilent Technologies). The RNA of the 50 samples had an average RNA integrity number (RIN) of 8.7 (ranging from 7.2 to 9.7). Libraries were prepared according to manufacturer&#39;s instructions (TruSeq RNA Sample Preparation kit) with an initial amount of 4 &mu;g of total RNA. Quality control of library templates was performed using a High Sensitivity DNA Assay kit (Agilent Technologies) on a Bioanalyzer (Agilent Technologies). Qubit quantification platform was used to normalize samples for the library preparation (Qubit 2.0 Fluorometer, Life Technologies). Libraries were sequenced via a paired-end chemistry on an Illumina HiSeq1000 platform with an average yield of &sim;6 Mb.</p>