Exosome microRNA Amplification and Profiling
Profile Exosome microRNAs from Serum
• Efficient purification and amplification of exosomal RNA
• Available for different sample types
• Human or Mouse 384 microRNA qPCR Profiler Assay included
RNAs present in patient body fluids are a rich and untapped source of disease-related biomarkers. The RNAs are stable in serum because they are encapsulated in circulating exosomes.
Exosomes are 40-100nm membrane vesicles secreted by most cell types in vivo and in vitro. They are found in blood, urine, amniotic fluid, malignant ascite fluids and contain distinct subsets of microRNAs depending upon the tumor or tissue from which they are secreted.
The Human and Mouse SeraMir Exosome RNA Amplification and Profiling Kits include everything needed to accurately and sensitively measure RNA from serum samples. Exosomes are efficiently isolated using ExoQuick Exosome Preciptation Solution, and exosomal RNA (exoRNA) is purified using a phenol-free lysis buffer and rapid spin columns.
The SeraMir exosome RNA amplification technology enables the 3‘ tailing and simultaneous tagging of both 5‘ and 3’ ends during cDNA synthesis, yielding cDNA ready for qPCR using the SeraMir Exosome RNA 384 microRNA qPCR Profiler Assay.
Primers for PCR amplification are included to make double-stranded cDNA that is ready to generate sense-strand exoRNA by T7 in vitro transcription. The resulting amplified exoRNA can be employed for microarray and NextGen Sequencing applications.
Representation between unamplified and amplified sequences is held constant for reproducible biomarker discovery and diagnostic development.
Choose the SeraMir Kit that’s right for you
Principle of the SeraMir Exosome RNA Amplification Kit
Exosomes are precipitated from biofluids, e.g. serum, with ExoQuick Exosome Precipitation Solution. Then exoRNAs are purified using a phenol-free lysis buffer and rapid spin columns. A poly(A) tail is added to the 3‘ end of exoRNAs in the presence of poly(A) polymerase. An oligo(dT) adaptor primer is used to initiate the subsequent reverse transcription reaction. When MMLV RT reaches the 5‘ end
of the RNA, it adds several non-template nucleotides, primarily deoxycytidines, to the 3‘ end of the newly synthesized first-strand cDNA. This oligo(dC) stretch base pairs to the complementary oligo(dG) sequence located at the 3‘ end of the 5’ Switch Adaptor. RT identifies the 5’ Switch Adaptor as an extra part of the RNA template and continues first-strand cDNA synthesis to the end of the oligonucleotide, thus incorporating the sequence into the 5‘ end of the cDNA.
The first-strand cDNA is ready for microRNA expression profiling. Alternatively, additional PCR amplification can be performed using the 5’ and 3’ adaptor primers, and the resulting double-stranded cDNA can then be further amplified by T7 in vitro transcription to yield exoRNA for microarray
or NextGen Sequencing applications.
Improved qPCR profiling of exosomal RNAs.
Serum RNA was prepared conventionally using TRIzol or with the SeraMir Exosome RNA Amplification Kit. Afterwards, expression of 380 human microRNAs was profiled using the SeraMir Exosome RNA 384 microRNA qPCR Profiler Assay, Cat# RA810A-1-SBI, see below.
The SeraMir RNA purification protocol combining phenol-free exosome lysis and small RNA binding columns isolates exoRNAs with much higher purity and reduced variability compared to TRIzol/phenol-based methods.
Three validated reference controls included on the SeraMir Profiling qPCR plate
• Perform Normalization with high confidence
• Use the geometric mean of all three reference gene Cts for relative qPCR
Related Links
User Manual
SeraMir Human 384 qPCR miRNA Primer List & Free Software
SeraMir Mouse 384 qPCR miRNA Primer List & Free Software
ExoQuick Exosome Precipitation Solution
Exosomal RNA Purification Kits
| Description | Cat# | Size | Price | ||
|---|---|---|---|---|---|
| Complete SeraMir Human Exosome RNA Amplification and Profiling Kit for Serum, Plasma, Tumor Ascites Fluid (contains cat-no. RA800A-1, RA805A-1, RA810A-1 components) | RA820A-1-SBI | 20 profiles | 1288 € | DETAILS | Add to Cart |
| Complete SeraMir Human Exosome RNA Amplification and Profiling Kit for Cell Culture Media, Urine, Spinal Fluid (contains cat-no. RA800A-1, RA805A-1, RA810A-1 and EXOTC50A-1 components) | RA820TC-1-SBI | 20 profiles | 1596 € | DETAILS | Add to Cart |
| SeraMir Human Exosome RNA 384 microRNA qPCR Profiler Primer Set | RA810A-1-SBI | 20 profiles | 550 € | DETAILS | Add to Cart |
| Complete SeraMir Mouse Exosome RNA Amplification and Profiling Kit for Serum, Plasma, Tumor Ascites Fluid (contains cat-no. RA800A-1, RA805A-1, RA811A-1 components) | RA821A-1-SBI | 20 profiles | 1288 € | DETAILS | Add to Cart |
| Complete SeraMir Mouse Exosome RNA Amplification and Profiling Kit for Cell Culture Media, Urine, Spinal Fluid (contains cat# RA800A-1, RA805A-1, RA810A-1 and EXOTC50A-1 components) | RA821TC-1-SBI | 20 profiles | 1596 € | DETAILS | Add to Cart |
| SeraMir Mouse Exosome RNA 384 microRNA qPCR Profiler Primer Set | RA811A-1-SBI | 20 profiles | 550 € | DETAILS | Add to Cart |
| SeraMir Exosome RNA Amplification Kit for Serum, Plasma, Tumor Ascites Fluid (contains cat.# R806A-1 plus reagents for tailing and tagging exoRNAs for cDNA synthesis and amplification) | RA800A-1-SBI | 20 rxn | 919 € | DETAILS | Add to Cart |
| SeraMir Exosome RNA Purification Kit for Serum, Plasma, Tumor Ascites Fluid (5ml Exoquick and 20 exoRNA Columns) | RA806A-1-SBI | 20 preps | 573 € | DETAILS | Add to Cart |
| SeraMir TC Exosome RNA Amplification Kit for Cell Culture Media, Urine, Spinal Fluid (contains cat.# R806TC-1 plus reagents for tailing and tagging exoRNAs for cDNA synthesis and amplification) | RA800TC-1-SBI | 10 rxn | 673 € | DETAILS | Add to Cart |
| SeraMir TC Exosome RNA Purification Kit for Cell Culture Media, Urine, Spinal Fluid (10ml ExoQuick-TC and 10 exoRNA columns) | RA806TC-1-SBI | 10 preps | 573 € | DETAILS | Add to Cart |
| SeraMir Control spike-in Small RNA and qPCR Assay | RA805A-1-SBI | 100 assays | 267 € | DETAILS | Add to Cart |
| SeraMir 3'-Universal Primer | RA800P-3-SBI | 1,2 ml for 10.000 reactions | 211 € | DETAILS | Add to Cart |
| Complete SeraMir Rat Exosome RNA Amplification and Profiling Kit (contains cat# RA800A-1, RA805A-1, RA812A-1 components) | RA822A-1-SBI | 20 profiles | 1288 € | DETAILS | Add to Cart |
| Complete SeraMir Rat Exosome RNA Amplification and Profiling Kit for Media and Urine (contains cat# RA800TC-1 with 50ml ExoQuick-TC, RA805A-1 and RA812A-1 components) | RA822TC-1-SBI | 20 profiles | 1596 € | DETAILS | Add to Cart |
| SeraMir Rat Exosome RNA 384 microRNA qPCR Profiler Primer Set | RA812A-1-SBI | 20 profiles | 550 € | DETAILS | Add to Cart |
