small RNA: EBOV-Minigenome with delta5-spacer transfected HEK293 cells, no VP30
From the small RNA samples CAP structures were removed from the small RNA samples, using RppH enzyme (NEB; analog to CAP-Clip Acid Pyrophosphatase). Afterwards Ribosomal RNA molecules were depleted, using an in-house developed protocol. Oligonucleotide adapters were ligated to the 5' and 3' ends of the RppH 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). The cDNA pool was size fractionated in the size range of 140 - 270 bp by PAA gel-electrophoresis.
SUB9183141
delta5_minusVP30_small_T2015_S14