CD Genomics offers tagRNA-seq for studying bacterial gene expression and regulation and has the potential to reveal new insights into the mechanisms underlying bacterial physiology and pathogenesis.

Overview of Our tagRNA-Seq Service

Our tagRNA-seq service is a cutting-edge method for studying bacterial gene expression and regulation. By leveraging short artificial RNA tags, we can distinguish between primary and processed transcripts and obtain a detailed picture of the bacterial RNA landscape. This method is particularly useful for identifying transcriptional start sites (TSSs) and processed start sites (PSSs) in bacteria, as well as detecting true antisense RNA transcripts (asRNAs) that are often difficult to detect with traditional RNA-seq methods. Our tagRNA-seq service provides a comprehensive and cost-effective solution for studying bacterial gene expression and regulation, enabling researchers to uncover new insights into the mechanisms underlying bacterial physiology and pathogenesis. With our team of expert scientists and state-of-the-art equipment, we offer reliable and high-quality tagRNA-seq data to support your research needs.

Library Construction of tagRNA-Seq

The RNA is then treated with tobacco acid pyrophosphatase (TAP) to remove the 5' triphosphate group from primary transcripts, leaving only the 5' monophosphate group on processed transcripts. Next, RNA adapters with unique tags are ligated to the 5' ends of the RNA molecules, with different tags for primary and processed RNAs. This allows for the differentiation of primary and processed transcripts during sequencing. The tagged RNA is then reverse-transcribed into cDNA and amplified using PCR. The resulting cDNA library is then subjected to high-throughput sequencing using Illumina or other next-generation sequencing platforms. The sequencing data can be analyzed using various bioinformatics tools to identify TSSs, PSSs, and asRNAs, and to obtain a comprehensive picture of the bacterial transcriptome. The library construction of tagRNA-seq requires careful optimization and quality control at each step to ensure reliable and reproducible results.

Project Workflow

Data analysis

• Genome-wide localization of polyadenylation sites
• Detection of TSS and PSS
• Gene expression level analysis
• Differential Gene Expression Analysis

Deliverable: FastQ, BAM, QC report, overview of TSS and PSS, snapshot of a processed RNA landscape, Custom bioinformatics analysis.

References:

1. Innocenti, Nicolas, et al. Detection and quantitative estimation of spurious double stranded DNA formation during reverse transcription in bacteria using tagRNA-seq. RNA biology 12.9 (2015): 1067-1069.
2. Innocenti, Nicolas, et al. Whole-genome mapping of 5′ RNA ends in bacteria by tagged sequencing: a comprehensive view in Enterococcus faecalis. Rna 21.5 (2015): 1018-1030.