Journal: Frontiers in Microbiology
Article Title: An Optimized Method for Extracting Bacterial RNA from Mouse Skin Tissue Colonized by Mycobacterium ulcerans
Figure Lengend Snippet: Evaluation of the coextraction method (Method 1) (A) Image of the electrophoresis gel for three independent RNA samples extracted from mice infected with M. ulcerans . L, RNA ladder. (B) Electropherogram: example of sample RNA derived from lane 1. (C) Table with RNA concentrations (ng/μl), indicator of purity (ratio A260/A280), integrity of RNA samples (RQI, RNA Quality Indicator), and total quantity (μg) of RNA extracted with Method 1. (D) Electropherogram: example of sample RNA before and after MicrobEnrich (ME) treatment. The 18S and 28S (mouse), and 16S and 23S (bacterial) rRNA bands are indicated in red and green, respectively.
Article Snippet: Total RNA was purified with the RNeasy Midi kit (Qiagen), with DNase treatment, according to the manufacturer’s protocol, and eluted in 100 μl RNase- and DNase-free water. (iii) DNase treatment: Contaminating DNA was removed by retreating the RNA with DNase, for 45 min at 37°C, with the TURBO DNA-free kit (Ambion), according to the manufacturer’s protocol. (iv) Eukaryotic RNA removal: To reduce the levels of contaminating host RNA from the samples, MICROBEnrich kit (Ambion) was used according to the manufacturer’s protocol.
Techniques: Electrophoresis, Mouse Assay, Infection, Derivative Assay