title: GSM9264366: EDL933 25 micromolar norepinephrine, biological replicate 3; Escherichia coli; ncRNA-Seq
library_name: GSM9264366
library_construction_protocol: The isolation of small RNAs from bacterial cells was performed using the PureLink miRNA Isolation Kit with slight modifications. Cells were lysed using TRIzol (Sigma-Aldrich), followed by chloroform treatment and centrifugation to separate the RNA-containing aqueous phase. Total RNAs were passed through Spin Cartridges with different ethanol concentrations to selectively bind and separate large and small RNAs. Finally, the small RNAs were eluted in RNase-free water after washing and a final centrifugation step. cDNA libraries were prepared using 75 ng of small RNAs as the input, following a variation of the AQRNA-seq library constuction protocol (PMCID: PMC8355021, PMID: 38372376) with two modifications. First, the use of a high-processivity reverse transcriptase replaced the need for AlkB demethylation. Second, an ExoI digestion was included immediately after the reverse transcription step to mitigate primer-dimer formation and eliminate the need for gel purification. Constructed libraries were subjected to quality assessment using Fragment Analyzer and qPCR.
identifiers.namespace: GEO
identifiers.id: GSM9264366_r1