GSM9652613: Fungal_mouse_24h_rep3_1; Candida albicans; RNA-Seq
24 h and 48 h after intravenous injection of Candida albicans into mice, the kidneys were harvested, and RNA was extracted from them. Candida albicans cultured in vitro was used as the control group. 1.1 Sample collection and preparation  1.1.1 RNA quantification and qualification  RNA degradation and contamination, especially DNA contamination, was monitored on 1.5% agarose gels.RNA concentration and purity was mesured using the advanced molecular biology equipment to ensure the use of qualified samples for transcriptome sequencing.  1.1.2 Library preparation for lncRNA-Seq  A total amount of 1.5 μg RNA per sample was used as input material for rRNA removal using the Ribo-Zero rRNA Removal Kit (Epicentre, Madison, WI, USA). Sequencing libraries were generated using NEBNextR UltraTM Directional RNA Library Prep Kit for IlluminaR (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. Briefly, Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer （ 5X）. First strand cDNA was synthesized using random hexamer primer and Reverse Transcriptase. Second-strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. In order to select insert fragments of preferentially 150~200 bp in length, the library fragments were purified with AMPure XP Beads (Beckman Coulter, Beverly, USA). Then 3 μl USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index(X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 and qPCR.  1.1.3 Clustering and sequencing  The clustering of the index-coded samples was performed on acBot Cluster Generation System using TruSeq PE Cluster Kitv3-cBot-HS(Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced onan Illumina platform and paired-end reads were generated.