small RNA: EBOV-Minigenome with NheI-Mutation transfected HEK293 cells, no VP30
CAP structures were removed from the small RNA samples, using CAP-Clip Acid Pyrophosphatase (Cellscript). Afterwards Ribosomal RNA molecules were depleted, using the Ribo-Zero rRNA Removal Kit GOLD for Human-Mouse-Rat (Illumina). Oligonucleotide adapters were ligated to the 5' and 3' ends of the CAP-Clip Acid Pyrophosphatase treated and rRNA-depleted small RNA samples. First-strand cDNA synthesis was performed using M-MLV reverse transcriptase and the 3 adapter as primer. The resulting cDNAs were 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. The sRNA pool was size fractionated in the size rage of 140 - 270 bp by PAA gel-electrophoresis.
SUB9183141
NheI_minusVP30_small_T1842_S3