GSM9363417: leuS-0.0125-rep1; Pseudomonas aeruginosa; RNA-Seq
50ul of treated cells were mixed with 25ul 3X RNAgem Blue Buffer, incubated at 75C for 10mins and flashfrozen. RNA was extracted using the Direct-zol RNA Miniprep Plus Kit. cDNA libraries were generated using a modified version of the RNAtag-seq protocol. 500ng of RNA was fragmented, depleted of genomic DNA, dephosphorylated, and ligated to DNA adapters. Barcoded RNAs were pooled and depleted of rRNA. Pools were converted to cDNA using reverse transcriptase template switching with PCR amplification of the Illumina P5 and P7 sequences. cDNA libraries were sequenced on Illumina Nextseq 500 platorm for paired end reads