Total RNA was extracted from Tetrahymena cells using the RNeasy Protect Cell minikit (Qiagen), as described (TetraFGD, http://tfgd.ihb.ac.cn/index/smphelp ). Polyadenylated transcripts were enriched using Sera-Mag magnetic oligo-dT beads (GE). First strand-specific libraries were constructed using the Illumina TruSeq Stranded mRNA sample preparation kit (RS-122-2101). Small RNA was enriched by a mirVana™ miRNA isolation kit (Ambion). Small RNA libraries were constructed using the Illumina TruSeq Small RNA Sample Prep kit (RS-200-0012). Sequencing was performed using an Illumina HiSeq-2500 sequencer. Analysis of polyadenylated transcripts was performed as described previously (Xiong et al. 2012 (link); Feng et al. 2017 (link)). Analysis of small RNA was performed as described (Schoeberl et al. 2012 (link)).
Protocol detail
Tetrahymena Transcriptome and Small RNA Profiling
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Corresponding Organization :
Other organizations : Ocean University of China, University of Michigan–Ann Arbor, Institute of Hydrobiology, Chinese Academy of Sciences, Qingdao National Laboratory for Marine Science and Technology, University of Utah, Cornell University, J. Craig Venter Institute
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Variable analysis
independent variables
- Method of total RNA extraction (RNeasy Protect Cell minikit)
- Method of polyadenylated transcript enrichment (Sera-Mag magnetic oligo-dT beads)
- Method of first strand-specific library construction (Illumina TruSeq Stranded mRNA sample preparation kit)
- Method of small RNA enrichment (mirVana miRNA isolation kit)
- Method of small RNA library construction (Illumina TruSeq Small RNA Sample Prep kit)
- Sequencing platform (Illumina HiSeq-2500)
- Total RNA extracted from Tetrahymena cells
- Polyadenylated transcripts
- Small RNA
- Tetrahymena cells
- Not specified
- Not specified
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