RNAs present in patient body fluids are a rich and untapped source of disease-related biomarkers and stable in serum because they are encapsulated in circulating exosomes.
The Human, Mouse and Rat 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.
Isolate biofluid exosomes and purify exoRNAs
Tail exoRNAs and synthesize double-tagged cDNA
Use the SeraMir spike-in RNA control in a qPCR assay to control for exoRNA recovery, tailing, and cDNA synthesis
Better qPCR profiling with SeraMir
Serum RNA prepared by the SeraMir Kit delivers more reliable, reproducible qPCR profiles than when the RNA is isolated using conventional Trizol methods. Profiling of 380 Human microRNAs across the SeraMir 384 Profiler. The phenol-free exosome lysis step coupled to the small RNA binding columns isolates exoRNAs with much higher purity than Trizol/Phenol based methods. The SeraMir exoRNAs are compatible with downstream polyadenylation and reverse trancription reactions for amplification and accurate qPCR profiling.
In addition, it is possible to apply real-time qPCR directly from exosomes without any RNA purification with the ExoCt RT-qPCR System.