Figure 2 | Scientific Reports

Figure 2

From: Cytometry meets next-generation sequencing – RNA-Seq of sorted subpopulations reveals regional replication and iron-triggered prophage induction in Corynebacterium glutamicum

Figure 2

Experimental workflow for the transcriptome analysis of bacterial populations after cell sorting. Cultures were analyzed by flow cytometry and sorted by the means of the fluorescent reporter signal. One million cells were sorted and immediately treated with an RNA stabilization agent (RNAlater or RNAprotect). Subsequently, cells were concentrated on a filter plate, flash frozen in liquid nitrogen and stored at −80 °C. Prior to RNA extraction, the cells were treated with lysozyme and mutanolysine. The quality of RNA was determined as RIN value (>7 for samples used for sequencing). Extracted RNA of appropriate quality was then used for cDNA library preparation and sequencing.

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