EVeryRNA™ EV RNA Purification System with SmartSEC Single (10 reactions)
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EVeryRNA™ EV RNA Purification System with SmartSEC Single (10 reactions). Cat# EVery110SS. Supplier: SBI System Biosciences</
Combine the contaminant-trapping enhanced SEC-based purification of SmartSEC Single EV isolation technology with the total RNA isolation capabilities of EVeryRNA EV RNA Purification technology for a complete solution for your EV RNA biomarker workflows.
The EVeryRNA EV RNA Purification System overcomes many of the challenges faced when isolating RNA from extracellular vesicles (EVs), most notable is the ability to capture total EV RNA, including small RNAs. EVeryRNA is effective even with low amounts of input RNA and is capable of delivering high yields of highly pure RNA. Because the RNA elutes in a small sample volume, thus generating a highly concentrated prep, you can increase the amount of RNA used in a single downstream reaction for better data coverage quality.
- Move quickly and confidently with exoRNA isolation that's high-yield and complete in <30 minutes
- Find what others miss when you capture every RNA with EVeryRNA
- Achieve phenol-level yields with a safer column-based method
- Get more RNA for each downstream reaction with EVeryRNA's small-volume elutions
- Ensure delivery of highly pure RNA by using the included DNaseI
- Isolate RNA from EVs for a full range of downstream applications, such as RNA-seq and miRNA profiling
- Maximize productivity with SmartSEC Single bundled with EVeryRNA
The EVeryRNA EV RNA Purification System with SmartSEC Single comes with sufficient reagents to perform 10 EV isolation reactions followed by EV RNA purification. EVeryRNA technology is available bundled with several of SBI's powerful EV isolation technologies as well as a cDNA Synthesis and Pre-amplification Kit (Table 1).
Table 1. EVeryRNA EV RNA Purification Products
How It Works
Isolate EVs with SmartSEC Single
The SmartSEC Single workflow is fast and easy. Simply apply 100 – 250 µL of cleared serum or plasma with additional column buffer or up to 4 mL of other biofluids directly to the pre-washed column, incubate, and centrifuge to elute the EVs.
Purify EV RNA with EVeryRNA
The EVeryRNA EV RNA Purification System delivers high yields of highly concentrated RNA from already isolated EVs. The column-based workflow is easy to implement and can be completed in less than 30 minutes (Figure 2).
To understand how well SmartSEC Single performs compared to a competitor’s q SEC columns, we isolated EVs from 250 µL of human serum using both SmartSEC Single and q SEC columns and analyzed 1 µg of protein equivalent from each isolation method using fluorescent nanoparticle tracking analysis (fNTA, Figure 3). EVs isolated using SmartSEC Single show a higher number of particles per mg of protein (Figure 3) than EVs isolated using the q SEC columns.
EVeryRNA captures EVerything
To demonstrate the ability of the EVeryRNA EV RNA Purification System to capture the full range of RNAs, we used the kit to isolate RNA from 10,000 cells (Figure 4, lane 1), from EVs that were isolated from 250 µL of serum using SmartSEC Single (Figure 4, lane 2), and from buffer spiked with 0.1 pmol of Cel-miR-39 (Figure 4, lane 3). The high quality of the isolated RNA can be seen in lane 1, where the RNA integrity number (RIN) is 9.9 and the 28S/18S RNA ratio is 1.5. The multiple bands in lane 2 demonstrate that EVeryRNA captures RNAs of different lengths—EVerything—from EVs with no apparent bias or size preference. The strong signal from the spiked-in miRNA in lane 3 demonstrates the good recovery of even small RNAs.
EVeryRNA works well with SmartSEC Single, yielding broad and unbiased size distribution of RNAs (Figure 5).
EVeryRNA delivers similar amounts of RNA as phenol-based methods
To demonstrate the excellent RNA yields obtained with EVeryRNA, we isolated EVs from 250 µL of serum using SmartSEC Single, spiked in 0.1 pmol of Cel-miR-39, and used both EVeryRNA and a phenol-based kit to isolate RNA. The isolated RNA was reverse transcribed using the EVeryRNA cDNA Synthesis & Pre-amplification Kit and the copy number of Cel-miR-39 measured (Figure 6). The EVeryRNA EV RNA Purification System delivered similar levels of Cel-miR-39 as the phenol-based method.
EVeryRNA efficiently isolates mRNA
We used the EVeryRNA EV RNA isolation kit to isolate mRNA from cells overexpressing eGFP (Figure 7). Robust levels of eGFP mRNA are recovered when cells are overexpressing eGFP.
miRNA isolated using EVeryRNA can be used for miRNA profiling and RNA-seq
We isolated EVs from 250 µL of serum using SmartSEC Single and used the EVeryRNA EV RNA Purification System and EVeryRNA cDNA Synthesis & Pre-amplification System to isolate and reverse transcribe EV RNAs for miRNA profiling using the SeraMir Human Exosome RNA Profiling Plate. We were able to detect a number of miRNAs both with amplification (96 miRNAs) and without amplification (16 miRNAs, with 14 overlapping with the miRNAs detected with amplification, Figure 8).
We were also able to show robust, successful RNA-seq runs using RNA isolated from EVs with EVeryRNA (Table 2). All three EV isolation methods tested generated high-quality RNA-seq data.
Table 2. Successful RNA-seq with EVeryRNA-isolated EV RNA