title: GSM3834647: ToxKO_Tim3- Day8_rep3; Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted using the Agencourt RNAdvance Cell v2 kit (A47942, Beckman Coulter). RNA integrity number (RIN) and yield were assessed using RNA 6000 Pico Kit (5067-1513, Agilent). cDNA synthesis and PCR amplification using 1 ng of total RNA from each sample was performed using SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (634891, Takara/Clontech). After cDNA synthesis, each sample was subjected to 12 cycles of PCR amplification. The generated amplicons were assessed and their concentration was determined with the use of Agilent High Sensitivity DNA Kit (5067-4626, Agilent). 150 pg of the resulting amplified cDNA were used for library preparation with the Illumina Nextera XT DNA Library reagents (FC-131-1024, Illumina). After PCR amplification of the fragmented libraries, the samples were purified with (0.6x) Agencourt AMPure XP beads and eluted in 10 µl of molecular grade water. The quality of the resulting library was assessed with the use of Agilent High Sensitivity DNA Kit (5067-4626, Agilent). The library quantification was performed based on the Illumina recommendations (SY-930-1010, Illumina) with the use of KAPA SYBR FAST qPCR Master Mix (KK4600, Kapa Biosystems).
attributes.tag: GEO Accession
attributes.value: GSM3834647