title: Serum RNA sample from cervical cancer women
organism: Homo sapiens
description: RNA was extracted from the serum of two groups ( healthy and cervical cancer patients). The total RNA quantity and purity were checked. The library preparation was performed using NEB Next Multiplex Small RNA library prep kit for Illumina according to the provided protocol. Further, PCR amplification and indexing were performed using Long Amp Taq 2X Master Mix. The PCR products were column purified using the Qiagen QIA quick PCR purification kit and eluted in nuclease-free water. The column-purified samples were loaded into a NOVEX 6% TBE gels and electrophoresed and stained with ethidium bromide solution. Two fractions corresponding to around145 base pair products were excised using a sterile scalpel and transferred into a 1.5mL tube containing 1 mL of DNA Gel Elution Buffer and vortexed at 350 rpm on a Thermal mixer for overnight to dissolve the gel completely. The library was purified using ice-cold ethanol and the size and concentrations were checked using an Agilent 4150 Tapestation followed by Qubit 3.0 fluorometer. The fraction of 140-150 base pair containing miRNAs of interest was taken for the sequencing in HiSeq 4000 System.