Prokaryotic mRNA-seq

Prokaryotic mRNA Sequencing is a transcriptome-profiling method for quantifying gene expression in prokaryotic organisms. Following depletion of abundant ribosomal RNA, the remaining RNA is sequenced to characterise coding and non-coding transcripts, compare expression profiles across conditions and investigate regulatory responses.

Prokaryotic mRNA-seq workflow

Applications of Prokaryotic mRNA-seq

Technical specifications of Prokaryotic mRNA-seq

• Library preparation: removal of ribosomal RNA from the sample
• Sequencing mode: 2 x 150 bp (PE150), stranded
• Sequencers: Illumina NovaSeq platform
• Recommended amount of data per sample:
  • 2 Gb per sample
  • Other configurations are also available depending on the client's needs.
• Data quality: Q30 ≥ 85%
• Recommended Sample Requirements - DNA/RNA:
  • Sample type: total RNA
  • Amount: > 1500 ng
  • Concentration (NanoDrop): >100 ng/μL
  • Volume: >15 μL
  • Purity (OD260/280): 1.8 - 2.2
  • RIN: >7 
• MINIMUM Sample Requirements - DNA/RNA:
  • Sample type: total RNA
  • Amount: > 1,000 ng
  • Concentration (Qubit): >50 ng/μL
  • Volume: >10 μL
  • Purity (OD260/280): 1.7 - 2.5
  • RIN: >6.5
  • 5.0 ≥ 28S/18S ≥ 1.0

RNA isolation from the supplied samples is also available. Details here.

Prokaryotic mRNA-seq bioinformatics analysis

The advanced bioinformatics analysis The Prokaryotic mRNA-seq package includes, among other analyses:

• Quality control of sequencing data
• Gene expression analysis
• Differential-expression analysis
• Functional annotation and enrichment analysis
• sRNA prediction and annotation
• Transcript structure analysis