GSE108190
Next Generation Sequencing Facilitates Quantitative Analysis of original and mutant E. coli K-12 MG1655 Transcriptomes
The goals of this study are to use Next-generation sequencing (NGS)to detect bacterial mRNA profiles of original E. coli K-12 MG1655 and fluoxetine induced E. coli mutants in response to 100 mg/L fluoxetine for 8 h, in triplicate, using Illumina HiSeq 2500.The NGS QC toolkit (version 2.3.3) was used to treat the raw sequence reads to trim the 3'-end residual adaptors and primers, and the ambiguous characters in the reads were removed. Then, the sequence reads consisting of at least 85% bases were progressively trimmed at the 3'-ends until a quality value = 20 were kept. Downstream analyses were performed using the generated clean reads of no shorter than 75 bp. The clean reads of each sample were aligned to the E. coli reference genome (NC_000913) using SeqAlto (version 0.5). Cufflinks (version 2.2.1) was used to calculate the strand-specific coverage for each gene, and to analyze the differential expression in triplicate bacterial cell cultures. The statistical analyses and visualization were conducted using CummeRbund package in R (http://compbio.mit.edu/cummeRbund/). Gene expression was calculated as fragments per kilobase of a gene per million mapped reads (FPKM, a normalized value generated from the frequency of detection and the length of a given gene. Overall design: Bacterial mRNA profiles of original E. coli K-12 MG1655 and fluoxetine induced E. coli mutants in response to 100 mg/L fluoxetine for 8 h, in triplicate, using Illumina HiSeq 2500