Ideal for use with small volumes of cell free fluids
- Saves money - does not require expensive equipment (ultracentrifuge and rotors)
- Saves time – exosome and microvesicle capture and RNA extraction can be completed in minutes instead of hours – days
- Allows fractionation of different types of microparticles
- Even difficult cell free fluids can be processed
- Maximizes RNA recovery
- Ideal for miRNA and mRNA studies of microvesicles and exosomes
The ExoMir™-MINI Kit is designed for filtration-based capture of exosomes and other microparticles from small volumes of cell-free biofluids, and for extraction of RNA from the captured particles. An appropriate range of biofluid volumes to use with the kit are from about 0.1 mL – 2 mL, although volumes outside this range are not necessarily incompatible with the kit. Traditional methods for recovering exosomes and other microparticles involve centrifuging biofluid samples at increasing centrifugal force, to sequentially pellet the larger and smaller particles. To recover exosomes, ultracentrifugation of samples at relative centrifugal force of at least 100,000 x g for several hours or more is generally required.
The ExoMir-MINI Kit uses an alternative approach for concentrating microparticles, in which they are captured by passing the samples over syringe filters. The filters are then flushed with an RNA extraction reagent to lyse the captured particles and release their contents. In the standard procedure, samples are passed over 2 filters connected in series, where the top filter has a larger pore size of approximately 200 nanometers to effectively capture larger particles such as apoptotic bodies and microvesicles, and the bottom filter has a smaller pore size of approximately 50 nanometers for capturing the smaller particles including exosomes. Examples of cell-free fluids that can be processed in this way include blood serum, plasma, urine, saliva, and eukaryotic cell culture media (“conditioned media”). After the sample has passed through the filters, they are disconnected and separately flushed with BiooPure™-MP to lyse the captured particles and release their contents. BiooPure-MP is a single-phase RNA extraction reagent containing guanidinium thiocyanate and phenol, which has been optimized to provide maximal recovery of the low-mass amounts of RNA in microparticles. Recovery of RNA is further improved by using the inert co-precipitant (linear acrylamide) included in the kit.
The ExoMir-MINI Kit contains sufficient components to treat and fractionate 10 samples of cell-free fluids, using 2 filters for each sample, and to extract RNA from both filters as well as from the flow-through filtrate.
The shelf life of the ExoMir-MINI Kit is 12 months when stored properly.