Gel Blot Hybridization for Viroids. 2022

Pedro Serra
Instituto de BiologĂ­a Molecular y Celular de Plantas (UPV-CSIC), Consejo Superior de Investigaciones CientĂ­ficas-Universitat Politecnica de Valencia, Valencia, Spain. pedseral@upvnet.upv.es.

Northern blot analysis reveals information about RNA identity, size, and abundance. This technique has become an essential tool for the knowledge developed about viroids and also an excellent method for viroid detection. Here we describe the methodology of a Northern blot based in polyacrylamide gel electrophoresis under denaturing conditions, hybridized with a viroid full-length riboprobe labeled with chemiluminescence. Viroid detection with this approach entails positive signals, specific migration, and the differentiation of their circular and linear forms.

UI MeSH Term Description Entries
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D014772 Viroids A group of pathogens comprising the smallest known agents of infectious disease. They are unencapsulated and are capable of replicating autonomously in susceptible cells. Positively identified viroids composed of single-stranded RNA have been isolated from higher plants, but the existence of DNA viroids pathogenic to animals is suspected. Viroid
D015152 Blotting, Northern Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES. Northern Blotting,Blot, Northern,Northern Blot,Blots, Northern,Blottings, Northern,Northern Blots,Northern Blottings

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