title: GSM8371093: ptth120BC2,L3-96h-AEL,rep1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: The samples were transferred to 1.7 ml centrifuge tube with a stainless-steel ball and 500ml Trizol. A0 was collected on the CO2 and transferred to a 1.7 ml centrifuge tube with a stainless-steel ball and 500ml Trizol. Tissuelyzer was used to smash the flies. About 15 pupa or flies were used in a biological replicate. Three biological replicates for each time points per genotype(wild type and Ptth mutants). RNA was sent to NOVOGENE for library and sequence after the total RNA was treated with DNase by the Turbo DNA-free kit (Invitrogen by Thermo Fisher Scitific, AM1907). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the cDNA was synthesized using random hexamer primers.  The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.