SeraMir Exosome RNA Amplification

Exosome RNA Amplification and Profiling

SeraMir

Overview:

Amplify Exosome RNAs for Profiling
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 40–100 nm membrane vesicles secreted by most cell types in vivo and in vitro. Exosomes 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 SeraMir kit includes everything needed to accurately and sensitively measure RNAs from serum samples. Exosomes are efficiently isolated using SBI’s ExoQuick solution, and the exoRNAs are purified using a phenol-free lysis buffer and rapid spin columns. The SeraMir kit enables the 3‘ tailing and simultaneous tagging of both 5‘ and 3’ ends during cDNA synthesis - ready for qPCR. Primers for PCR amplification are included to make double-stranded cDNA that is ready to generate sense-strand exoRNAs using for T7 IVT. Use the amplified exoRNAs for microarrays and NextGen sequencing applications.

Go from serum sample to amplified exoRNAs in one day
SeraMir Overview

Technical Details:

How the SeraMir Kit Works

• Precipitate exosomes from patient biofluids easily with ExoQuick
• Purify exoRNAs with SeraMir columns
• Tail and tag all exoRNAs for qPCR, cDNA amplification and T7 IVT
• No RNA ligase used - avoid adaptor concatemer artifacts
• Amplify exoRNAs for Microarrays and NextGen Sequencing

Precipitate serum exosomes and purify exoRNAs

Precipitate serum exosomes and purify exoRNAs

Tail exoRNAs and synthesize double-tagged cDNA

Tail exoRNAs and synthesize double-tagged cDNA

qPCR Profiling:

Better qPCR Profiling with SeraMir

qPCR_comparison

Serum RNA prepared by conventional Trizol versus the SeraMir kit. Profiling of 380 Human microRNAs across the SeraMir 384 Profiler (cat# RA810A-1). 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.

SeraMir Spike-in RNA control

Use the SeraMir spike-in RNA control and qPCR assay to control for exoRNA recovery, tailing and cDNA synthesis.

SpikeIN_qPCR

384 SeraMir microRNA Profiler cat# RA810A-1

Measure the top 380 microRNAs by qPCR across 20 exoRNA samples tagged using the SeraMir kit. The qPCR array also includes triplicate reactions for the RNA spike-in control.