Prokaryotic RNA sequencing profiles the transcriptome through a stranded RNA library, delivering a more accurate estimation of transcript expression.
Application
- Profiling transcripts in prokaryotes
- Predicting operon, TSS, promoter, and UTR
- Exploring RNA regulation mechanism through antisense transcripts prediction and sRNA analysis
Benefits
- Novogene adopted rRNA depletion methods for the construction of prokaryotic RNA libraries. Effective depletion methods were adopted for a variety of species.
- Besides gene expression quantification, multiple gene structure and gene function analysis are included in the standard analysis. Customized analyses can also be devised for prokaryotic RNA sequencing.
Specifications: RNA Sample Requirements
| Library Type | Sample Type | Amount | RNA Integrity Number (Agilent 2100) | Purity (NanoDrop) |
| Prokaryotic RNA Library | Total RNA | ≥ 500 ng | ≥ 6.0, smooth base line | A260/280 ≥ 2.0;
A260/230 ≥ 2.0 |
Specifications:Sequencing and Analysis
| Sequencing Platform | Illumina NovaSeq 6000 Sequencing System |
| Read Length | Paired-end 150bp |
| Recommended Data Amount | ≥ 2Gb raw data per sample for the species with reference genome |
| Content of Data Analysis | Data Quality Control
Alignment to Reference Genome Gene Expression Analysis Novel Transcript Prediction Gene expression quantification & Differential expression analysis Enrichment Analysis Operon, Promoter and TSS/TTS Prediction SD Sequence Prediction Rho-independent Terminator Sequence Prediction sRNA Secondary Structure Prediction sRNA Targeted Gene Prediction |
Project Workflow
The first step of the project workflow includes sample quality control (Sample QC) to ensure that your samples meet the criteria of the RNA-Seq technique. Then, an appropriate library is constructed according to your target organism and subsequently tested for its quality (Library QC). Next, a paired-end 150 bp sequencing strategy is used to sequence the samples, and the resulting data goes through quality data control (Data QC) to guarantee the quality of the resulting data. Finally, bioinformatic analyses are performed and publication-ready results are provided. The following flowsheet elaborates the step-by-step protocol.
