title: GSM2109567: WT_2 h_RNA-seq; Tetrahymena thermophila; RNA-Seq
library_construction_protocol: Cells for each of the samples with the amount of approximately 1×107 (cells) for each samples were washed once with 1×PBS (centrifuge at 450×g for 2min) and spun down to pellet followed by immediate freezing in liquid nitrogen.The total RNA of the freezed cell samples were then extracted by Trizol lysis followed by chloroform and isopropanol treatment. After extract total RNA and treated with DNase I, Oligo(dT) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA are fragmented. Then  cDNA  is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for the  PCR  amplification. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time  PCR  System are used in quantification and qualification of the sample library. Then the library is sequenced using Illumina HiSeq 4000 .
attributes.tag: GEO Accession
attributes.value: GSM2109567