title: GSM9266699: Drosophila-head-WT-female-with-DMSO--1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: RNA was extracted from single head of 21 days old verging female w1118 / SIRT6 KO Drosophila fed with 100µM TDO2 inhibitor/ DMSO for 21 days.  RNA was extracted using BioTri RNA (Bio-Lab, catalogue number 959758027100), according to the manufacturer's manual followed by cleaning step using RNA Clean & Concentrator MagBead (Zymo Research, catalogue number R1081). Sample QC -Total RNA samples were quantified using Qubit 4.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked with 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA). Samples were initially treated with TURBO DNase (Thermo Fisher Scientific, Waltham, MA, USA) to remove DNA contaminants. ERCC RNA Spike-In Mix (Cat: #4456740) from ThermoFisher Scientific, was added to normalized total RNA prior to library preparation following manufacturer's protocol. The next steps included performing rRNA depletion using QIAGEN FastSelect rRNA HMR or Fly Kit (Qiagen, Germantown, MD, USA), which was conducted following the manufacturer's protocol. RNA sequencing libraries were constructed with the NEBNext Ultra II RNA Library Preparation Kit for Illumina by following the manufacturer's recommendations. Briefly, enriched RNAs are fragmented for 15 minutes at 94 °C. First strand and second strand cDNA are subsequently synthesized. cDNA fragments are end repaired and adenylated at 3'ends, and universal adapters are ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200 (Agilent Technologies, Palo Alto, CA, USA), and quantified using Qubit 4.0 Fluorometer (ThermoFisher Scientific, Waltham, MA, USA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). Azenta US, Inc. 2910 Fortune Circle West, Suite E, Indianapolis, IN 46241 | Tel: (858)527 - 7080 . The sequencing libraries were multiplexed and clustered on the flowcell. After clustering, the flowcell was loaded on the Illumina NovaSeq instrument according to manufacturer's instructions. The samples were sequenced using a 2x150 Pair-End (PE) configuration.
identifiers.namespace: GEO
identifiers.id: GSM9266699_r1