GSM3577224
GSM3577224: sample4_BDM_L. pneumophila corby_48h; Homo sapiens; miRNA-Seq
RNA was extracted from cells by phenol-chloroform precipitation Samples with sufficient RNA quality were used for Illumina library preparation according to the manufacturer´s instructions. Briefly, 3' and 5' adapters were ligated to 1 µg total RNA with T4 RNA ligase (Epicentre). Following the reverse transcription of the RNA into cDNA, PCR amplification was performed. To distinguish the prepared samples, different index primers were added. Afterwards, samples with unique indices were pooled into two libraries and cDNA was loaded on a PAA-gel. The fraction with a size of 145 – 160 bp was cut from the gel to enrich for small RNAs. This fraction contained the cDNA transcribed from mature miRNAs generated from 22 nt RNA fragments. Following elution of the cDNA, the final libraries were concentrated by ethanol precipitation. Finally, prepared libraries were validated by a Bioanalyzer run and sent for sequencing.
GEO
GSM3577224
GEO Accession
GSM3577224