NRI Nutritional Biochemistry Core-Kannapolis

The NRI Nutritional Biochemistry Core-Kannapolis provides a variety of cutting edge nutritional biochemical analyses for choline and its metabolites, as well as other cellular metabolism and cell analysis techniques.

PCR Single Nucleotide Polymorphism (SNP) Genotyping for One-Carbon Metabolism
Purified DNA from blood or tissues is analyzed by PCR for single nucleotide polymorphisms (SNPs) of your specification. In most cases, TaqMan® SNP Genotyping Assays will be used to detect SNPs and real time PCR carried out on an Eppendorf Mastercycler epgradient Realplex4 (Eppendorf). We specialize in genotyping for one carbon metabolism SNPs: PEMT rs1235718, CHDH rs12676 and CHDH rs9001, however other SNPs can be analyzed.

Mitochondrial Function Test (Seahorse Platform)
This assay allows for real time measurement of basal respiration, ATP turnover, maximal respiration, spare capacity and proton leak all in the same well of cells. Respiration, that is the oxygen consumption rate (OCR), and the extracellular acidification rate (ECAR) are measured simultaneously in each well of the plate by special solid state sensor probes. The XF24 Extracellular Flux Analyzer (Seahorse Bioscience) runs plates with 20 experimental wells and 4 control wells. We supply the special assay plates for you to grow your cells, and we will run the XF Mito Stress Kit (Seahorse Bioscience) for you.

Cellular Glycolysis Test (Seahorse Platform)
This assay yields real time cellular measurements of glycolysis, glycolytic capacity and glycolytic reserve, all in the same well of cells. Glycolysis is measured by the extracellular acidification rate, or ECAR. The oxygen consumption rate (OCR) is also measured simultaneously in same wells. ECAR and OCR are measured by special solid state sensor probes. Our XF24 Extracellular Flux Analyzer (Seahorse Bioscience) runs plates with 20 experimental wells and 4 control wells. We supply the special assay plates for you to grow your cells, and we will run the XF Glycolysis Stress Test Kit (Seahorse Bioscience) for you.

DIY Seahorse XF24 Platform
This is an hourly rate for use of the Seahorse XF24 platform. This assumes prior training or experience with the XF24 Analyzer and associated software. Users are expected to supply their own plates, reagents, calibrant and medium.

Seahorse XF24 Training
Training on the Seahorse XF24 Analyzer (Seahorse Biosciences) is an 8 hour, hands-on course where you will learn how to setup a protocol and run a plate of cells using the Mito Stress Test Kit. First you will measure basal respiration or the oxygen consumption rate (OCR), and the extracellular acidification rate (ECAR) which are measured simultaneously in each well of the plate by special solid state sensor probes. The trainee will learn how to process the data and how to design future experiments in their cell of choice.

Zeiss Axio Imager Fluorescence Microscope with Camera (DIY)
The Zeiss Axio Imager A1 microscope and camera is available for hourly rental. This microscope has brightfield, darkfield, differential phase interference contrast (DIC), polarization contrast (PO) and fluorescence capabilities. Available filter sets for DAPI, FITC and rhodamine detection. AxioVision 4.8.2 software available on attached pc.

Genomic DNA Extraction
DNA is extracted using a column-based method such as Qiagen’s QIAamp DNA Blood Mini Kit (Qiagen). Column methods are efficient, automatable, and have less chance of yielding samples with contaminants that could inhibit downstream applications. DNA will be dissolved in 10 mM Tris-HCL pH 8.0, 1mM EDTA (TE) (equivalent to Qiagen’s buffer AE). We use a NanoDrop ND-1000 Spectrophotometer to check the DNA concentration and purity.

Folate Assay (RBC or Plasma)
Folate is measured in red blood cells (RBC) or plasma samples, using a microbiological method.

Folate Assay (Food or Tissue)
Folate is measured in food or tissue samples, using a microbiological method.

Homocysteine Assay (ELISA)
Homocysteine is measured in plasma or serum samples by an enzyme linked immunoadsorbent assay (ELISA) (IBL International).

Choline Metabolites-Choline, Phosphocholine, Glycerophosphocholine, Betaine, TMAO, Methionine, Creatinine, Phosphatidylcholine, and Sphingomyelin
This assay is designed to measure choline and related metabolites listed above by liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Plasma, urine, breast milk, food or tissue samples may be submitted.

Aqueous Choline Metabolites-Choline Phosphocholine, Glycerophosphocholine, Betaine, TMAO, Methionine and Creatinine
This assay is designed to measure choline and related metabolites listed above by liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Plasma, serum urine, breast milk, tissue samples or liquid food may be submitted.

Organic Choline Metabolites-Phosphatidylcholine, and Sphingomyelin
This assay is designed to measure the metabolites listed above by liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Plasma, urine, breast milk or tissue samples may be submitted.

Dimethyl-glycine, Choline and Betaine
This assay is designed to measure the metabolites listed above by liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Plasma, urine, food or tissue samples may be submitted.

TMAO, Creatinine, Choline and Betaine
This assay is designed to measure trimethylamineoxide, creatinine, choline and betaine by liquid chromatography-stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM/MS). Plasma, urine, breast milk or tissue samples may be submitted.

Fatty Acids
This assay is designed to measure fatty acids by thin layer chromatography/gas chromatography (TLC/GC). Plasma, breast milk, food or tissue samples may be submitted.

Phospholipid Fatty Acid Assays
This assay is designed to measure phospholipid fatty acids by thin layer chromatography/gas chromatography (TLC/GC). Plasma, breast milk, food or tissue samples may be submitted.


Core Personnel

zeisel_steve_2012Steven H. Zeisel, MD, PhD, Center Director
Kenan Distinguished Professor
Departments of Nutrition & Pediatrics
T: (919) 843-4731
E: steven_zeisel@unc.edu

Steve Orena, Core Manager
UNC Nutrition Research Institute
Zeisel Lab

T: (704) 250-5041
E: orena@email.unc.edu


Service Request Form

To request NRI nutritional biochemistry services, please complete the online service request form. For more information about core services, please contact Steve Orena, Core Manager.