This protocol describes how to isolate exosomal RNA from cell culture supernatant using the Exoquick Seramir tissue culture kit.
Method Article
Isolation of exosomal RNA from cell culture supernatant using the System Biosciences Exoquick Seramir tissue culture kit
https://doi.org/10.1038/protex.2017.085
This work is licensed under a CC BY-NC 3.0 License
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This protocol describes how to isolate exosomal RNA from cell culture supernatant using the Exoquick Seramir tissue culture kit.
exRNA
extracellular vesicles
exosomes
RNA isolation
Exoquick
SeraMir
Extracellular RNAs (exRNAs) have been found to play an important role in intercellular communication in the body. exRNAs are present in biofluids, in complexes with lipoproteins and ribonucleoproteins, and in extracellular vesicles such as exosomes and microvesicles. Further study and characterization of exRNAs and their carriers could lead to identification of new biomarkers and have potential for development of novel therapeutics.
Exoquick Seramir tissue culture kit (Systems Biosciences, RA800-TC1)
100% ethanol (VWR, 89125-186)
Microfuge
Microfuge tubes, 1.5 mL
1 This volume was selected to match that of other kits to enable fair comparisons. As little as 10 μL RNase-free water can be used for elution if a higher RNA concentration is required, but the yield will be reduced by approximately 20%. Do not elute with less than 10 μL RNase-free water, as the spin column membrane will not be sufficiently hydrated. The dead volume of the RNeasy MinElute spin column is 2 μL: elution with 14 μL RNase-free water results in a 12 μL eluate.
2 Centrifuging at a low speed first helps the solvent wet the surface of the membrane prior to the full speed centrifuging step. This results in a better yield / RNA recovery from the membrane.
This protocol was modified from the manufacturer's instructions for the Exoquick Seramir kit.
KD, BS and SD were supported by NIH grant UH3TR000901. SS and LL were supported by U01 HL126494 and UH3 TR000906. IY and TP were supported by UH3 TR000884.
The authors declare no competing financial interests.
This protocol has been posted on Protocol Exchange, an open repository of community-contributed protocols sponsored by Nature Portfolio. These protocols are posted directly on the Protocol Exchange by authors and are made freely available to the scientific community for use and comment.
posted
You are reading this latest protocol version