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外泌体RNA合成与预扩增

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  • 外泌体RNA合成与预扩增
    SeraMir Exosome RNA Amplification Kit Everything you need to accurately and sensitively measure exosomal RNAs from serum, plasma, or ascites fluid.Description
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    Complete SeraMir Exosome RNA Amplification kit (5 mL ExoQuick, 20 exoRNA columns, cDNA synthesis and amplification components)


    20 Reactions
    RA800A-1
    • Reliable, reproducible exoRNA isolation from serum, plasma, or ascites fluid
    • Sensitive detection via exoRNA amplification
    • Excellent preparation method for qPCR
    • Can be used to amplify sense-strand exoRNAs for microarray construction

    With cargo that reflects the makeup of their parent cells and their easy isolation, researchers are increasingly turning to exosomes as a source of disease-related biomarkers. To simplify and standardize the isolation of RNA from exosomes, SBI has developed the SeraMir family of products.

    The SeraMir Exosome RNA Amplification Kit includes everything you need to accurately and sensitively measure RNAs from serum samples—ExoQuick for fast and efficient exosome isolation, a phenol-free lysis buffer and rapid spin columns for exoRNA isolation, and reagents for 3’ tailing and simultaneous tagging of both 5’ and 3’ ends during cDNA synthesis so you can go straight to qPCR.

    In addition, primers for PCR amplification are included to make double-stranded cDNA via T7 IVT. These amplified exoRNAs can be used to construct microarrays or for next generation sequencing applications.

    Choose the SeraMir Kit that’s right for you

    Cat. # Name Kit includes
        ExoQuick or ExoQuick-TC SeraMir RNA columns and reagents SeraMir cDNA synthesis and amplification reagents 384-well plate miRNAs for human, mouse, or rat
    RA800A-1 Complete SeraMir Exosome RNA Amplification Kit        
    RA800TC-1 Complete SeraMir Exosome RNA Amplification Kit for Media and Urine        
    RA806A-1 SeraMir Exosome RNA Purification Kit        
    RA806TC-1 SeraMir Exosome RNA Purification Kit for Media and Urine        
    RA808A-1 SeraMir Exosome RNA Purification Column Kit        
    RA820A-1 Human Complete SeraMir Exosome RNA Amplification and Profiling Kit        
    RA820TC-1 Human Complete SeraMir Exosome RNA Amplification and Profiling Kit for Media and Urine        
    RA821A-1 Mouse Complete SeraMir Exosome RNA Amplification and Profiling Kit        
    RA821TC-1 Mouse Complete SeraMir Exosome RNA Amplification and Profiling Kit for Media and Urine        
    RA822A-1 Rat Complete SeraMir Exosome RNA Amplification and Profiling Kit        
    RA822TC-1 Rat Complete SeraMir Exosome RNA Amplification and Profiling Kit for Media and Urine        

     

     

    How It Works

    Go from sample to amplified exoRNAs in a single day

    • Isolate exosomes from patient biofluids with the included ExoQuick reagent
    • Purify exoRNAs with SeraMir columns
    • Tail and tag all exoRNAs for qPCR
    • Perform second strand synthesis to generate cDNAs for amplification and T7 IVT

    The amplified exoRNAs are ready for use in microarrays or NGS. Because RNA ligase is not used, you can avoid complicating adaptor concatemer artifacts.

    Isolate serum 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.

     

    Supporting Data

    Better qPCR profiling with SeraMir

     

    Figure 1. 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.

     

    Figure 2. Serum exoRNAs prepared using SeraMir deliver excellent performance in microarray studies. Samples from a pooled normal serum preparation and from a male caucasian (age 73) with adenocarcinoma of the colon were used in this study. Exosomes were precipitated from 250 µL of serum using the SeraMir Exosome RNA Amplification Kit. The T7-amplified “sense” exoRNAs were then used for direct labeling analyses on LC Sciences miRBase ver.16 array chips (performed in triplicate). The exoRNAs were hybridized across 1,214 different microRNAs on the probe set.

    Of the 1,214 microRNAs analyzed, 79 microRNAs showed a signal intensity >32. Within this set of 79, there was a clear colon versus normal “signature set” of 40 microRNAs that could discriminate normal from colon cancer serum samples with a p-value < 0.01. The identities of the microRNAs found in this study have been masked while further investigation continues.

     


    Product Documentation
    • User Manual: SeraMir Exosome RNA Amplification Kits
    Citations
    • Zhang, R, et al. (2017) Serum long non coding RNA MALAT-1 protected by exosomes is up-regulated and promotes cell proliferation and migration in non-small cell lung cancer. Biochem. Biophys. Res. Commun.. 2017 Jun 13;. PM ID: 28623135
    • Huleihel, L, et al. (2017) Matrix bound nanovesicles recapitulate extracellular matrix effects on macrophage phenotype. Tissue Eng Part A. 2017 Jun 4;. PM ID: 28580875
    • Peng, Y, Huleihel, L & Cardenes, N. (2017) Different MiroRNA Expression In MSC-Derived Exosomes: IPF Patients And Age-Matched Normal Individuals. Conference. ;. Link: Conference
    • Protocol, IIEI. (2017) List of Components. Product. ;. Link: Product
    • Tsukita, S, et al. (2017) MicroRNAs 106b and 222 Improve Hyperglycemia in a Mouse Model of Insulin-Deficient Diabetes via Pancreatic β-Cell Proliferation. EBioMedicine. 2017 Feb 1; 15:163-172. PM ID: 27974246
    • Josson, S, Gururajan, M & WKChung, L. (2017) miRNA Characterization from the Extracellular Vesicles. bio-protocol. ; 7(4). Link: bio-protocol
    • Val, S, et al. (2017) Purification and characterization of microRNAs within middle ear fluid exosomes: implication in otitis media pathophysiology. Pediatr. Res.. 2017 Apr 5;. PM ID: 28157838
    • Sueta, A, et al. (2017) Abstract P1-02-13: Differential expression of exosomal miRNAs between breast cancer patients with recurrence and no-recurrence. Abstract. ;. Link: Abstract
    • Selmaj, I, et al. (2017) Global Exosome Transcriptome Profiling Reveals Biomarkers for Multiple Sclerosis. Ann. Neurol.. 2017 Apr 15;. PM ID: 28411393
    • Zhou, X, et al. (2017) Characterization of mouse serum exosomal small RNA content: The origins and their roles in modulating inflammatory response. Oncotarget. 2017 Jun 27; 8(26):42712-42727. PM ID: 28514744
    • Hubal, MJ, et al. (2017) Circulating adipocyte-derived exosomal MicroRNAs associated with decreased insulin resistance after gastric bypass. Obesity (Silver Spring). 2017 Jan 1; 25(1):102-110. PM ID: 27883272
    • Nakano, T, et al. (2017) Hepatic miR-301a as a Liver Transplant Rejection Biomarker? And Its Role for Interleukin-6 Production in Hepatocytes. OMICS. 2017 Jan 1; 21(1):55-66. PM ID: 28271982
    • Holliday, LS, et al. (2017) Exosomes: novel regulators of bone remodelling and potential therapeutic agents for orthodontics. Orthod Craniofac Res. 2017 Jun 1;:95-99. PM ID: 28643924
    • Faust, A, et al. (2017) Urinary bladder extracellular matrix hydrogels and matrix-bound vesicles differentially regulate central nervous system neuron viability and axon growth and branching. J Biomater Appl. 2017 Apr 1; 31(9):1277-1295. PM ID: 28447547
    • Koyama, S, et al. (2017) Dynamic changes of serum microRNA-122-5p through therapeutic courses indicates amelioration of acute liver injury accompanied by acute cardiac decompensation. ESC Heart Fail. 2017 May 1; 4(2):112-121. PM ID: 28451447
    • Reza, AM, et al. (2016) Human adipose mesenchymal stem cell-derived exosomal-miRNAs are critical factors for inducing anti-proliferation signalling to A2780 and SKOV-3 ovarian cancer cells. Sci Rep. 2016 Dec 8; 6:38498. PM ID: 27929108
    • Hsu, CY, et al. (2016) Synthetic Steroid Hormones Regulated Cell Proliferation Through MicroRNA-34a-5p in Human Ovarian Endometrioma. Biol. Reprod.. 2016 Mar 1; 94(3):60. PM ID: 26819477
    • Gutierrez, MJ, et al. (2016) Airway Secretory microRNAome Changes during Rhinovirus Infection in Early Childhood. PLoS ONE. 2016 Sep 20; 11(9):e0162244. PM ID: 27643599
    • Huang, Y, et al. (2016) 血清中外泌体及外泌体 RNA 提取方法的比较. 中华检验医学杂志. 2016 Jul 20; 39:427-432. Link: 中华检验医学杂志
    • Gomez, D & Sempere, L. (2016) Characterization of Exosomal microRNAs in Pancreatic Cancer. Thesis. ;. Link: Thesis
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