GSM4594402: Human_L861-01-T03; Homo sapiens; miRNA-Seq
All samples in this study were enrolled from Gansu Provincial Hospital between October 2018 and March 2019. The cohort from this study was comprised of patients with lung adenocarcinoma and healthy peoples. We collected the plasma sample from research subjects that had been confirmed with tissue pathology. Also, all patients were not accepted any therapeutic strategies, such as surgery, chemoradiotherapy, targeted treatment and immunotherapy, and had no any complications or other history of cancer.Five milliliters of peripheral venous blood were collected in from each object with ethylenediaminetetraacetic acid (EDTA) anticoagulant tube, and then centrifuged (TD5G, Shanghai) at 1000 rpm for 10 minutes at room temperature immediately, followed by a 10 minutes high-speed centrifugation (Biofuge Frsco, Heraeus) at 12,000 rpm at 4°C, and finally, removed cell debris. The supernatant plasma was stored at -80 °C.A total amount of 2.5ng RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNextR UltraTM small RNA Sample Library Prep Kit for IlluminaR (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. Briefly, first of all, ligated the 3′ SR Adaptor. Mixed 3′ SR Adaptor for Illumina, RNA and Nuclease-Free Water, mixture system after incubation for 2 minutes at 70 degrees in a preheated thermal cycler. Tube was transfer to ice. Then, add 3′ Ligation Reaction Buffer (2X) and 3′ Ligation Enzyme Mix ligate the 3′ SR Adaptor. Incubated for 1 hour at 25°C in a thermal cycler. To prevent adaptor-dimer formation, the SR RT Primer hybridizes to the excess of 3′ SR Adaptor (that remains free after the 3′ ligation reaction) and transforms the single stranded DNA adaptor into a double-stranded DNA (dsDNA) molecule. dsDNAs are not substrates for ligation mediated. The second, ligated the 5′ SR Adaptor. Then, reverse transcription synthetic first chain. PCR amplification and Size Selection. PAGE gel was used to electrophoresis fragment screening purposes, rubber cutting recycling as the pieces get small RNA libraries. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system.
GEO Accession
GSM4594402