The RBL Microbiology Unit is a collaborative research service component of the Regional Biocontainment Laboratory at Duke (housed in GHRB). The Unit maintains quality-controlled secure inventories of approved BSL2, BSL3, and Select Agent bacterial pathogens for RBL projects and collaborating investigators. The Microbiology Unit has expertise in Chlamydia, and TB research program and helps the collaborators with study design, protocol optimization in-vitro and in-vivo. The core provides services such as IVIS live animal imaging in ABSL2/3, aerosol-delivered infection, macrophage-mycobacterium infection, extraction of mycobacteria from human samples, qRT-PCR, transcervical chlamydia infection (mice), and Inclusion Forming Unit (IFU) assay of chlamydia.
Manager: Uma Shankar Gautam, PhD
Minimum Inhibitory Concentration assay, small molecule inhibitors, and compound testing (EC50 and IC50 determination)
Screening of chemical compounds and small molecule inhibitors in cell-based assays under BSL2 and BSL3 settings.
Colony forming unit assay and Quality controlled bacterial stocks propagation: BSL2, BSL3, and select agents
Determination of bacteria in liquid cultures and aerosol samples, freezing bacterial stocks for stable, long-term storage. Our freezing methods allow recovering cultures with maximum viability.
Mycobacteria isolation from human tuberculosis specimens (sputum), stock propagation, genomic DNA, and RNA isolation
Processing of sputum from human TB patients and isolating mycobacteria, propagating clinical isolates of mycobacteria in liquid cultures, and isolation of sequencing grade genomic DNA and RNA from mycobacteria.
THP1 cell line and primary macrophage infection with mycobacteria
Cell culture work for BSL2/BSL3-specific agents: Isolation of Bone Marrow-Derived macrophages, differentiation, and activation, of mammalian cells (e.g., THP1 cells), and infection with mycobacteria, CFU counts, sequencing grade RNA isolation and RNAi-based experiments in infected macrophages.
Transient transfection of mammalian cells and antibody purification
Transient transfection of Expi293 cells and purifying the antibodies.
Endotoxin testing and coordinating research collaborations by providing recommendations to facilitate decision-making for the following or similar samples but not limited to:
- DNA/ RNA/ Nucleic acid encapsulated in Lipid Nano-particles
- Virus preps
- Chemical compounds etc.
Microplate reader-based bacterial quantification controlled with Gen5 data analysis software interface. Micro-volume nucleic acid quantification with Take3Trio plates (reads 48 samples at once). Gene amplification and qRT-PCR assay (96 well and 384 wells).
Bacterial and fungal challenge model in mice
The microbiology core has established bacterial doses for infecting animals with Mycobacterium tuberculosis, Yersinia pestis, and sexually transmitted diseases e.g., Chlamydia spp., and fungal agents e.g., Coccidioides immitis, etc. We provide customized full or partial support to meet your microbiology lab and animal model needs, including aerobiology (see Animal Support).
Project-specific support/ other
The Microbiology core will also consider developing new emerging technologies or other services that would be valuable for investigators for BSL2 and BSL3 agent-specific work: Mycobacterium tuberculosis, Yersinia pestis, Chlamydia, macrophages, antibody purification from mammalian cells, and targeted dose delivery of bacterial agents in animal model.
Contact RBL Microbiology Unit Manager with needs.
Assay Request & Sample Submission Tool
Step 1: Download and complete the appropriate sample submission manifest
More Coming Soon!
Step 2: Complete the Assay Request Form
Current approved rates for Duke Investigators can be seen in CoreResearch@Duke (Core D-0045)
CoreResearch@Duke can also be used to request an ESTIMATE for a grant or contract proposal
Service Requests and Estimates
Internal Duke users:
Make a Service REQUEST (Instructions)
External Non-Profit Institution users:
- A signed Research Analysis Quote and Agreement (RAQA) and Purchase Order (PO) will be needed from your institution
- Contact Unit manager for a price quote to get the process started
Non-Duke For-Profit Entity users:
- Duke Shared Resources can only provide services to For-Profit Entities under a signed Sponsored Research Agreement (SRA)
- The collaborative research project must have a defined research question, scope of work, budget, and timeline.
- Contact Unit manager to get the process started.
Directions to the RBL and Parking Information (contact us)
How to acknowledge work done in our facility:
Please acknowledge our facility in any publications/presentations: “____work was performed in the Microbiology Unit of the Regional Biocontainment Laboratory at Duke, which received partial support for construction from the National Institutes of Health, National Institute of Allergy and Infectious Diseases (UC6-AI058607; G20-AI167200).”