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Method Article
Roopali Gandhi
Department of Neurology, Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
Corresponding Author
Louise C. Laurent
Department of Reproductive Medicine, University of California, San Diego, La Jolla, CA, USA
Corresponding Author
Anil K. Sood
Departments of Gynecologic Oncology and Reproductive Medicine and of Cancer Biology and Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center (MDACC), Houston, TX, USA
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol describes how to isolate total extracellular RNA from serum or plasma using the Norgen BioTek Plasma/Serum circulating and exosomal RNA purification mini kit. It supercedes an earlier protocol (http://dx.doi.org/10.1038/protex.2015.116). The new protocol is based on our experience vetting the original protocol in multiple labs. The major change is an increase in input volume of biofluid from 200 µL to 500 µL. We found that the larger input volume led to more reproducible amounts of RNA isolation across multiple experiments. We have also included footnotes to explain changes we made to manufacturers' protocols, and observations that were made while carrying out the protocols.
Keywords
exRNA, extracellular vesicles, exosomes, serum, plasma, blood, Norgen BioTek
The authors declare no competing financial interests.
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Method Article
Roopali Gandhi
Department of Neurology, Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
Corresponding Author
Louise C. Laurent
Department of Reproductive Medicine, University of California, San Diego, La Jolla, CA, USA
Corresponding Author
Anil K. Sood
Departments of Gynecologic Oncology and Reproductive Medicine and of Cancer Biology and Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center (MDACC), Houston, TX, USA
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 16 Jun, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol describes how to isolate total extracellular RNA from serum or plasma using the Norgen BioTek Plasma/Serum circulating and exosomal RNA purification mini kit. It supercedes an earlier protocol (http://dx.doi.org/10.1038/protex.2015.116). The new protocol is based on our experience vetting the original protocol in multiple labs. The major change is an increase in input volume of biofluid from 200 µL to 500 µL. We found that the larger input volume led to more reproducible amounts of RNA isolation across multiple experiments. We have also included footnotes to explain changes we made to manufacturers' protocols, and observations that were made while carrying out the protocols.
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