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Title:
Small RNA Library Construction from Minute Biological Samples | SpringerLink
Description:
Increasingly, the discovery and characterization of small regulatory RNAs from a variety of organisms have all required deep-sequencing methodologies. However, the crux to successful deep-sequencing analysis depends upon optimal construction of a cDNA library...
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Keywords {🔍}

pubmed, small, article, scholar, google, rnas, cas, rna, methods, central, drosophila, sequencing, protocol, library, lau, analysis, piwiinteracting, micrornas, genes, biol, privacy, cookies, content, information, publish, minute, biology, mol, siomi, research, search, construction, samples, igloi, regulatory, cdna, endogenous, pirnas, access, chapter, bartel, science, cell, dev, download, springer, usd, personal, data, log,

Topics {✒️}

month download article/chapter xenopus piwi-interacting rnas dicer-dependent small rnas required deep-sequencing methodologies high-throughput sequencing platform piwi-interacting rnas gung-wei chirn & nelson small rna-seq methods small rna profiling cdna library sequencing privacy choices/manage cookies cdna library compatible device instant download small endogenous rnas enriched follicle cells small rna sequencing small rna preparations small rna cloning drosophila piwi proteins small rnas isolated small regulatory rnas regulatory small rnas small expressed rnas plant small rnas author information authors gung-wei chirn deep sequencing technology single cell analysis drosophila rna methyltransferase searle scholar large-scale sequencing european economic area antisense oligonucleotide depletion polyacrylamide gel system probable regulatory roles mediates 2′-o-methylation babiarz je substantially expanded set orr-weaver tl van den engh structural rna fragments minute biological samples journal finder publish conditions privacy policy ruby jg previously annotated genes differentially expressed genes successful library accepting optional cookies modifies germline pirnas

Schema {🗺️}

ScholarlyArticle:
      headline:Small RNA Library Construction from Minute Biological Samples
      pageEnd:136
      pageStart:123
      image:https://media.springernature.com/w153/springer-static/cover/book/978-1-62703-694-8.jpg
      genre:
         Springer Protocols
      isPartOf:
         name:PIWI-Interacting RNAs
         isbn:
            978-1-62703-694-8
            978-1-62703-693-1
         type:Book
      publisher:
         name:Humana Press
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:Jessica A. Matts
            affiliation:
                  name:Brandeis University
                  address:
                     name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Yuliya Sytnikova
            affiliation:
                  name:Brandeis University
                  address:
                     name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Gung-wei Chirn
            affiliation:
                  name:Brandeis University
                  address:
                     name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Gabor L. Igloi
            affiliation:
                  name:University of Freiburg
                  address:
                     name:Institute of Biology, University of Freiburg, Freiburg, Germany
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Nelson C. Lau
            affiliation:
                  name:Brandeis University
                  address:
                     name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
                     type:PostalAddress
                  type:Organization
            type:Person
      keywords:Small RNAs, Illumina deep-sequencing, Library construction
      description:Increasingly, the discovery and characterization of small regulatory RNAs from a variety of organisms have all required deep-sequencing methodologies. However, the crux to successful deep-sequencing analysis depends upon optimal construction of a cDNA library compatible for the high-throughput sequencing platform. Challenges to small RNA library constructions arise when dealing with minute tissue samples because certain structural RNA fragments can dominate and mask the desired characterization of regulatory small RNAs like microRNAs (miRNAs), endogenous small interfering RNAs (endo-siRNAs), and Piwi-interacting RNAs (piRNAs). Here, we describe methods that improve the chances of constructing a successful library from small RNAs isolated from minute tissues such as enriched follicle cells from the Drosophila ovarium. Because the ribosomal RNA (rRNA) fragments are frequently the major contaminants in small RNA preparations from minute amounts of tissue, we demonstrate the utility of antisense oligonucleotide depletion and an acryloylaminophenylboronic acid (APB) polyacrylamide gel system for separating the abundant 2S rRNA in Drosophila from endo-siRNAs and piRNAs. Finally, our methodology generates libraries amenable to multiplex sequencing on the Illumina Hi-Seq platform.
      datePublished:2014
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Book:
      name:PIWI-Interacting RNAs
      isbn:
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         978-1-62703-693-1
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      name:Humana Press
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      name:Brandeis University
      address:
         name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
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         name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
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      name:Brandeis University
      address:
         name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
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      name:University of Freiburg
      address:
         name:Institute of Biology, University of Freiburg, Freiburg, Germany
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      name:Brandeis University
      address:
         name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
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Person:
      name:Jessica A. Matts
      affiliation:
            name:Brandeis University
            address:
               name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
               type:PostalAddress
            type:Organization
      name:Yuliya Sytnikova
      affiliation:
            name:Brandeis University
            address:
               name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
               type:PostalAddress
            type:Organization
      name:Gung-wei Chirn
      affiliation:
            name:Brandeis University
            address:
               name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
               type:PostalAddress
            type:Organization
      name:Gabor L. Igloi
      affiliation:
            name:University of Freiburg
            address:
               name:Institute of Biology, University of Freiburg, Freiburg, Germany
               type:PostalAddress
            type:Organization
      name:Nelson C. Lau
      affiliation:
            name:Brandeis University
            address:
               name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
               type:PostalAddress
            type:Organization
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      name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
      name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
      name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
      name:Institute of Biology, University of Freiburg, Freiburg, Germany
      name:Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, USA
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