Microbiome Core

About

The overall goal of BEES Sub-Core is to advance EHS research of CACHET members by providing ready access to state-of-the-art “-Omics” technologies and expertise across the two campuses, facilitating effective utilization of resources, enhancing training opportunities and providing subsidized rates for services to Center members.

Biomarkers of Exposure Effects and Susceptibility (BEES) Sub-Core

To accomplish this goal, the BEES Sub-Core leverages existing institutional resources at UofC and UIC in microbiome analysis, metabolomics, proteomics, genomics and bioinformatic services. To enable streamlined access to these varied services by CACHET members and provide advice on appropriate experimental design and resource utilization, each BEES Sub-Core has a Center-supported Facilitator who is the director of that institutional facility core and a subject-matter expert. In addition to providing guidance and priority access to CACHET members, they will provide training to CACHET investigators who wish to master these technologies, particularly young investigators and those new to the fields. The BEES Sub-Core uses the Office of Shared Research Facilities cores (OSRF) central web-based ordering system at UofC for services and subsidy/co-pay requests which enables efficient turnaround time for all CACHET members on both campuses. This system permits tracking of subsidized services and Core/Unit utilization in a centralized database for efficient metrics analysis of facility usage and outcomes.

Leadership

Ashley has been the Metabolomics Platform Director for the Host-Microbe Metabolomics Facility (HMMF) within the Duchossois Family Institute (DFI) at the University of Chicago since April 2021. Her career has focused on the study of host-microbe interactions mediated by microbial factors, specifically within inflammatory bowel disease (IBD). Following her Chemistry PhD work in untargeted metabolomics and analytical chemistry from Indiana University (mentor Dr. Erin E. Carlson), she completed her post-doctoral work at the University of Chicago with Dr. Dionysios A. Antonopoulos and Dr. Eugene B. Chang where she quantitatively and qualitatively explored peptides and metabolites from continuous microbial culture, IBD patients and murine model systems. She currently serves as a Scientific Advisor on the GIRF AB since 2020. In the DFI – HMMF, Ashley and the amazing HMMF team members (Jessica Little, PhD; Mary McMillin, BS; David Moran, MS, MPH; Michael Mullowney, PhD; Amber Rose, MS) are dedicated to providing targeted and untargeted metabolomics platforms to the host-microbe community to elucidate and define disease state-specific metabolite profiles for the discovery of early detection and prevention strategies as well as the discovery of novel treatments.

 

Anitha Sundararajan received her PhD from Montana State University in 2011 from the Center for Biofilm Engineering and the Department of Microbiology. For her dissertation, she studied two metabolically diverse organisms and how they sense and respond to different environmental stressors to optimize metabolism.  During this time, she developed and optimized several techniques to cultivate and harvest anaerobic biofilms (under microaerophilic conditions) with defined oxygen concentrations. For her post-doctoral training, she took up a position at the National Center for Genome Resources (NCGR), a sequencing service provider and a biotech company.  Here, she was trained on Illumina and Pacific Biosciences sequencing platforms and is well versed with library preparation workflows for DNA and RNA. Here at DFI she wishes to combine her microbiology background with her genomics experience and build a core that offers different services to support DFI’s mission, i.e. to investigate the gut microbiome and its implication(s) in a range of human diseases using -omics approaches.

 

 

Key Accomplishments

  • Trained Argonne Microbiome Core staff and developed specimen receipt protocols for EHS studies;
  •  Streamlined and refined microbiome sequencing assays, sample collection, processing procedures and bioinformatics pipelines for profiling gut and saliva microbiome in population studies;
  • Established protocols and collection procedures for air sample microbiomes from South Chicago households;
  • Provided microbiome profiling services to eight studies led by CACHET members, including three funded by the CACHET Pilot Project Program;
  • Recruited Dr. Eric Pamer to assume Microbiome Core Leader position upon Dr. Gilbert’s departure;
  • With establishment of the DFI, CACHET launched a collaborative arrangement for DFI to assume the role of CACHET Microbiome Core, with a focus on translational EHS research;
  • DFI purchased four new state-of-the-art MS systems dedicated to microbial metabolite profiling;
  • Recruited Dr. Chaubard of DFI as CACHET Facilitator for Metabolomics Core, particularly for EHS research;
  • Established collaborative arrangements to leverage pre-existing and high-functioning institutional genomics and bioinformatics core facilities at UofC to augment the new BEES Sub-Core services for CACHET members; key services for CACHET investigators will be provided at “in-house” service rates with subsidized co-pay support available for justified EHS research.

The DFI Microbiome Metagenomics Facility (MMF) provides bacterial 16S ribosomal RNA gene and shotgun metagenomic sequence analyses on intestinal contents and other samples containing complex microbial populations.  The MMF will work with investigators to analyze microbiota sequence data from experimental and clinical samples. Analysis tools are available to classify bacterial sequences down to the genus level (and to the species level when possible) to determine both community membership and structure. Analyses include graphic visualization of the bacterial composition of individual samples and determination of microbial population diversity within and between samples.

DFI MMF CACHET GRANT USER PRICING

Eppendorf EpMotion 5075

An automated liquid handling system that purifies, amplifies and splits the samples processed at the MMF.

 

Eppendorf EpMotion 5073

An automated liquid handling system that purifies, amplifies and splits the samples processed at the MMF.

 

QiaCube HT

Used for DNA extraction.

 

Qiagen TissueLyser II

The TissueLyser II simultaneously disrupts multiple biological samples through high-speed shaking in plastic tubes with stainless steel, tungsten carbide, or glass beads.

 

Maxwell RSC DNA Purification

The Maxwell® 16 Tissue DNA Purification Kit is used with the Maxwell® 16 Instrument to purify genomic DNA and RNA from up to 50mg of tissue samples in 30–40 minutes.

 

Agilent Tape Station 4200

The Agilent TapeStation system is an automated electrophoresis solution for the sample quality control of DNA and RNA samples. The system integrates an instrument, data processing software, reagents, and ScreenTape devices specific for DNA and RNA.

 

Illumina MiSeq

The Illumina MiSEQ accesses focused applications such as targeted resequencing, metagenomics, small genome sequencing, and targeted gene expression profiling. MiSeq reagents enable up to 15 Gb of output with 25 million sequencing reads and 2 × 300 bp read lengths.

 

Illumina NextSeq 1000/2000

The NextSeq 1000 and NextSeq 2000 Systems support a wide range of methods such as exome sequencing, target enrichment, single-cell profiling, transcriptome sequencing, and more.

 

Oxford Nanopore MinION

Oxford’s Nanopore reads the length of DNA or RNA presented to them — from short to ultra-long (longest >4 Mb). MinION supports whole genomes/exomes, metagenomics, targeted sequencing, whole transcriptome, smaller transcriptomes, and multiplexing for smaller samples.

1. Go to the Lab Link page: LabLink
2. Click “Request for a User ID”.
3. Fill out all required fields on form and press “Request User ID”.
4. Within 24 hours, you will receive an email notification when your ID request has been approved.
5. After approval, login to the LabLink page.
6. Go to the “Resource Materials” tab.
7. Download and fill out the template for the service you are requesting.
*You must fill out the template first or you will not be able to submit your project. Additionally, do not add any additional COLUMNS, COMMAS, or DASHES or your submission will not go through! *
8. Save your template somewhere convenient (i.e. your Desktop).
9. Go back to the LabLink page (Note: make sure you are on the “Projects” tab).
10. Press the “Create” button in the top right corner.
11. Fill out all required fields on the form and press “Continue”.
12. Press the “Browse Document” button and select your completed template from step 3.
13. Skip the “Upload Additional Documents” step.
14. Review all details of your project, and check all boxes in the disclaimer at the bottom of the form. Press
“Submit Project” when you are ready.

Contact Dr. Anitha Sundararajan for Experimental and Sample Requirements: ANITHAS@BSD.UCHICAGO.EDU

The DFI Metabolomics Core provides state-of-the-art MS-based metabolomic capabilities to identify, measure and characterize the metabolites produced/modified by microbes as effectors of microbe-microbe and host-microbe interactions that influence human health and disease. For example, butyrate, a microbiota-associated short-chain fatty acid (SCFA), was found to serve as an important immunomodulatory compound that contributes to respiratory tract infections (13, 14). This innovative metabolome approach to microbiome science, not typically available in microbiome facilities, provides a more comprehensive understanding of the health impacts that result from microbiome modifications due to disease, therapies or environmental exposures, the later a research topic of several Center members.

 

DFI HMMF CACHET GRANT USER PRICING

Agilent 8890/5977B GCMS with Chemical Ionization (CI)
A GCMS instrument dedicated to absolute and relative quantitation of short-chain fatty acids and a diversity of other metabolites using PFBBr derivitization.
 

Agilent 7890B/5977 GCMS with Chemical Ionization (CI)

A GCMS instrument dedicated to absolute and relative quantitation of short-chain fatty acids and a diversity of other metabolites using PFBBr derivitization.
 

Agilent 7890B/5977B GCMS with Electron Impact Ionization (EI)

A GCMS instrument used for both targeted and untargeted analysis of up to ~170 metabolites after TMS/MOX derivatization.
 

Agilent 1290 Infinity II / 6470 Triple Quadrupole (QqQ) LC-MS

Our Triple Quad provides high-precision MS/MS data for targeted quantitative analysis in our Indole (Tryptophan Pathway Metabolites / B Vitamins) panel.

 

Agilent 1290 Infinity II / 6546 Quadrupole Time-of-Flight (Q-TOF) LC-MS

The Agilent 6546 LC/QTOF is used for the absolute and relative quantitation of bile acids, custom targeted metabolomics, and high-resolution accurate mass (HRAM) untargeted metabolomics.

 

Thermo Scientific Vanquish UHPLC / Orbitrap IQ-X Tribrid LC-MS

The Thermo IQ-X mass spectrometer allows us to perform data-rich untargeted metabolomics and robust compound identification and structure elucidation of small molecules at very high resolution, while also being central to our custom targeted metabolomics platform.

 

Agilent 1290 Infinity II HPLC

The freestanding HPLC stack is used for UV detection, fractionation, and purification.

 

Gerstel MPS automated sample preparation system

The Gerstel MPS streamlines our sample preparation to enable the analysis of large sample sets.

1. Go to the Lab Link page: LabLink
2. Click “Request for a User ID”.
3. Fill out all required fields on form and press “Request User ID”.
4. Within 24 hours, you will receive an email notification when your ID request has been approved.
5. After approval, login to the LabLink page.
6. Go to the “Resource Materials” tab.
7. Download and fill out the template for the service you are requesting.
*You must fill out the template first or you will not be able to submit your project. Additionally, do not add any additional COLUMNS, COMMAS, or DASHES or your submission will not go through! *
8. Save your template somewhere convenient (i.e. your Desktop).
9. Go back to the LabLink page (Note: make sure you are on the “Projects” tab).
10. Press the “Create” button in the top right corner.
11. Fill out all required fields on the form and press “Continue”.
12. Press the “Browse Document” button and select your completed template from step 3.
13. Skip the “Upload Additional Documents” step.
14. Review all details of your project, and check all boxes in the disclaimer at the bottom of the form. Press
“Submit Project” when you are ready.

Contact Dr. Ashley Sidebottom for Experimental and Sample Requirements: ASIDEBOTTOM@BSD.UCHICAGO.EDU