title: small RNA: Mock Huh7 cells with EBOV leader Spike-in
design: Ribosomal RNA molecules were depleted, using the Ribo-Zero rRNA Removal Kit GOLD for Human-Mouse-Rat (Illumina). To the small RNA sample 2% of a vitro-transcript was added (65nt Spike-in). Afterwards the CAP structures were removed from the small RNA sample, using CAP-Clip Acid Pyrophosphatase (Cellscript). Oligonucleotide adapters were ligated to the 5' and 3' ends of the CAP-Clip Acid Pyrophosphatase treated and rRNA-depleted small RNA sample. First-strand cDNA synthesis was performed using M-MLV reverse transcriptase and the 3 adapter as primer. The resulting cDNA was PCR-amplified using a high fidelity DNA polymerase. The cDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics) and was analyzed by capillary electrophoresis.