sample_prep_urine.nan
NAN KB, Mario Uchimiya, John Glushka, Leandro I Ponce, Laura Morris, Arthur Edison, Saraa Al Jawad
Disclaimer
This protocol is developed and maintained by Network for Advanced NMR (NAN). The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to this protocol is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with this protocol, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.
Abstract
This is a modified protocol for NMR metabolomics for urine samples. This method was originally proposed by:
Before start
This protocol assumes that the original urine samples have >600 µL.
Steps
Day-1/1
2
Day-1/1
Dry the samples in a speed-vac concentrator
Thaw samples On ice or at 4°C
Centrifuge the samples at 4°C at 12000x g,0h 0m 0s for 0h 5m 0s
Dry the samples in a speed-vac concentrator
Add 60µL of the phosphate buffer to 540µL of the supernatant for each sample
- Use 1.5-mL Eppendorf tubes
Transfer the supernatant of each sample to a new tube
- Use 1.5-mL tubes
Vortex the samples at 4°C for 0h 2m 0s
Centrifuge the samples at 16000rcf for 0h 30m 0s
Centrifuge the samples at 4°C for 0h 0m 10s
Incubate the samples at -20°C for 0h 20m 0s
Transfer 590µL of the supernatant to an NMR tube for each sample
Vortex the samples for 0h 0m 10s
Add 600µL of 100% cold methanol to 300µL of samples On ice
- Use 1.5-mL tubes
Thaw samples On ice or at 4°C
