Home › Analytical Chem › Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform
Analytical Chem JoVE (Open Access) Citable · DOI

Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform

DOI: 10.3791/53460-v
What you'll learn
  • ✓ Extract RNA from infected tissue samples using standard protocols
  • ✓ Apply digital bar-coding platform to profile pathogen gene expression
  • ✓ Interpret gene expression results from microbial pathogens during infection
  • ✓ Evaluate sensitivity and reproducibility of pathogen profiling methods
Protocol

Biopharma Insights We developed a protocol that is fast, sensitive and reproducible for pathogen gene expression profiling during an infection.

Difficulty
advanced
Total time
~4–6 hours per infected tissue sample (RNA extraction through profiling)
Model organism
Infected tissue (protocol demonstrates Candida albicans infection)
Biosafety
BSL-2

Steps

1
Prepare reagents and instruments for profiling

Set up and calibrate all equipment, reagents, and digital bar-coding platform components required for downstream RNA extraction and gene expression analysis.

▶ 00:59
2
Extract RNA from infected tissue samples

Isolate total RNA from infected tissue using appropriate lysis and purification methods to obtain high-quality RNA for downstream profiling.

▶ 01:47
3
Perform gene expression profiling using digital bar-coding

Apply the digital bar-coding platform to quantify pathogen gene expression from extracted RNA with high sensitivity and reproducibility.

▶ 04:01
4
Analyze and interpret microbial gene expression results

Evaluate gene expression data from pathogenic microbes (e.g., C. albicans) to understand infection-stage-specific transcriptional responses.

▶ 06:55
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