RNA-Seq of wild type E. coli MG1655 versus those with a deletion in the hns gene (Histone-like protein H-NS). RNA-Seq of wild type E. coli MG1655 versus those with a deletion in the hns gene (Histone-like protein H-NS)

The aim of the experiment was to determine differences in RNA levels between wild-type E. coli and E. coli with an hns deletion. We used RNA-seq, but to control for variation in sample preparation and sequencing, we combined each E. coli RNA sample with a "spike-in” Salmonella RNA sample of known concentration. Libraries constructed in duplicate from strains grown with and without Salmonella spike-in to assess absolute genome-wide changes in transcription.This allowed us to normalise RNA levels between samples during data analysis.

本实验旨在探究野生型大肠杆菌（E. coli）与hns基因缺失型大肠杆菌之间的RNA水平差异。本研究采用RNA测序（RNA-seq）技术，为控制样本制备与测序环节的误差变异，将每份大肠杆菌RNA样品与已知浓度的沙门氏菌（Salmonella）spike-in RNA样品进行混合。针对添加与未添加沙门氏菌spike-in的大肠杆菌菌株样本，我们分别构建了两份重复测序文库，以评估全基因组范围内的绝对转录水平变化。该实验设计使得我们能够在数据分析阶段对不同样本间的RNA水平进行标准化校正。