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| 1 | Methods for the extraction and RNA profiling of exosomes显示文摘AIM:To develop protocols for isolation of exosomes and characterization of their RNA content.METHODS:Exosomes were extracted from He La cell culture media and human blood serum using the Total exosome isolation(from cell culture media)reagent,and Total exosome isolation(from serum)reagent respectively.Identity and purity of the exosomes was confirmed by Nanosight?analysis,electron microscopy,and Western blots for CD63 marker.Exosomal RNA cargo was recovered with the Total exosome RNA and protein isolation kit.Finally,RNA was profiled using Bioanalyzer and quantitative reverse transcriptionpolymerase chain reaction(q RT-PCR)methodology.RESULTS:Here we describe a novel approach for robust and scalable isolation of exosomes from cell culture media and serum,with subsequent isolation and analysis of RNA residing within these vesicles.The isolation procedure is completed in a fraction of the time,compared to the current standard protocols utilizing ultracentrifugation,and allows to recover fully intact exosomes in higher yields.Exosomes were found tocontain a very diverse RNA cargo,primarily short sequences 20-200 nt(such as mi RNA and fragments of m RNA),however longer RNA species were detected as well,including full-length 18S and 28S r RNA.CONCLUSION:We have successfully developed a set of reagents and a workflow allowing fast and efficient extraction of exosomes,followed by isolation of RNA and its analysis by q RT-PCR and other techniques. | Emily Zeringer Mu Li Tim Barta Jeoffrey Schageman Ketil Winther Pedersen Axl Neurauter Susan Magdaleno Robert Setterquist Alexander V Vlassov | 2013 | World Journal of Methodology2013，3，1： | 5 |
| 2 | Exosomes:current knowledge of their composition,biological functions,anddiagnostic and therapeutic potentials显示文摘 | Vlassov A V Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 3 | Exosome , currrent knowledge of their composition, biological function, and diagnostic and therapeutic potentials 显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 4 | Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials显示文摘 | Vlassov A V Magdaleno S Setterquist R | 2012 | Biochim Biopto s Acta2012，1820，7： | 1 |
| 5 | Exosomes: Current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials 显示文摘 | Vlassdv A V Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 6 | Exosomes: cur- rent knowledge of their composition, biological functions, and di- agnostic and therapeutic potentials 显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 7 | Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptc1^+/-p53^-/-mice显示文摘 | Romer JT KimuraH Magdaleno S | 2004 | Cancer Cell2004，6，3： | 1 |
| 8 | 2D-FFT Implementation on FPGA for Wavefront Phase Recovery from the CAFADIS Camera 显示文摘 | Rodriguez-Ramos Magdaleno J M | 2008 | SPIE Astronomical telescopes and instrumentation Marseille2008，7015，： | 1 |
| 9 | Pregnancy after bariatric surgery:a current view of maternal,obstetrical and perinatal challenges显示文摘 | Magdaleno R J Pereira B G Chaim E A | 2012 | Arch Gynecol Obstet2012，285，3： | 1 |
| 10 | Exosomes:Current knowledge of their composition,biological functions,and diagnostic and therapeutic potentials显示文摘 | Vlassov A Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 11 | Exosomes: Current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials显示文摘 | Alexander V. Vlassov Susan Magdaleno Robert Setterquist Rick Conrad | 2012 | BBA - General Subjects2012，，7： | 1 |
| 12 | Exosomes:current knowledge of their composition,biological functions,and diagnostic and therapeutic potentials显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 13 | Optimization of transfection conditions and analysis of siRNA potency using real-time PCR显示文摘 | Cheng A Magdaleno S Vlassov AV | | 0，，01： | 1 |
| 14 | Pilot describing function measurements in a multiloop task显示文摘 | STAPLEFORD R L MCRUER D T MAGDALENO R E | 1967 | IEEE Transactions on Human Factors in Electronics1967，8，2： | 1 |
| 15 | Actions of a proline ana- logue, L-thiazolidine-4-carboxyllic acid (T4C), on Trypanosoma cruzi显示文摘 | Magdaleno A Ahn I Y Paes I S | 2009 | PLoS One2009，4，2： | 1 |
| 16 | Exosomes: current knowledge of their composition,biological functions, and diagnostic and therapeutic potentials 显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Bio phys Aeta2012，1820，7： | 1 |
| 17 | Identification of the dominant ground handling characteristics of a navy jet trainer显示文摘 | Klyde D H Myers T T Magdaleno R E | 2002 | Journal of Guidance Control and Dynamics2002，25，3： | 1 |
| 18 | Early embryonic lethality in mice with targeted deletion of the CTP:phosphocholine cytidylyltransferase alpha gene (Pcyt1a)显示文摘 | Wang L Magdaleno S Tabas I | 2005 | Mol Cell Biol2005，25，8： | 1 |
| 19 | Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptcl ( +/-)p53(-/-) mice显示文摘 | Romer JT Kimura H Magdaleno S | 2004 | Cancer Cell2004，6，3： | 1 |
| 20 | Exosomes: current knowledge of their composition, biological functions, and diagnos- tic and therapeutic potentials显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Bioehim Biophys Acta2012，1820，7： | 1 |
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