title: GSM8241520: fur A18 F110 I3 R1; Escherichia coli str. K-12 substr. MG1655; RNA-Seq
library_construction_protocol: All samples from both rounds were prepared and collected in biological duplicates at UCSD. 3 ml of culture sampled from an overnight grown culture at an OD600 nm = 0.5 was added to 6 ml of Qiagen RNA-protect Bacteria Reagent after sample collection. This solution was then vortexed for 5 seconds, incubated at room temperature for 5 minutes, and then centrifuged. The supernatant was then removed and the cell pellet was stored at -80° C. The Zymo Research Quick-RNA MicroPrep Kit was used to extract RNA from the cell pellets per vendor protocol. On-columns DNase treatment was performed for 30 minutes at room temperature. Anti-rRNA DNA Oligo mix and Hybridase Thermostable RNase H31 was used to remove ribosomal RNA. Sequencing libraries were created using a Kapa Biosystems RNA HyperPrep per vendor protocol.