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    • Fulltext

       

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      Permanent link:
      https://www.ias.ac.in/article/fulltext/jbsc/039/05/0795-0804

    • Keywords

       

      Bacillus; isothermal probe; locked nucleic acid; oligonucleotide microarray

    • Abstract

       

      Oligonucleotide microarray has been one of the most powerful tools in the ‘Post-Genome Era’ for its high sensitivity, high throughput and parallel processing capability. To achieve high detection specificity, we fabricated an isothermal microarray using locked nucleic acid (LNA)-modified oligonucleotide probes, since LNA has demonstrated the advanced ability to enhance the binding affinity toward their complementary nucleotides. After designing the nucleotide sequences of these oligonucleotide probes for gram-positive bacilli of similar origin (Bacillus subtilis, Bacillus licheniformis, Bacillus pumilus, Bacillus megaterium and Bacillus circulans), we unified the melting temperatures of these oligonucleotide probes by modifying some nucleotides using LNA. Furthermore, we optimized the experimental procedures of hydrating microarray slides, blocking side surface as well as labelling the PCR products. Experimental results revealed that KOD Dash DNA polymerase could efficiently incorporate Cy3-dCTP into the PCR products, and the LNA-isothermal oligonucleotide microarray were able to distinguish the bacilli of similar origin with a high degree of accuracy and specificity under the optimized experimental condition.

    • Author Affiliations

       

      Jing Yan1 Ying Yuan1 Runqing Mu2 Hong Shang2 Yifu Guan1

      1. Key Laboratory of Medical Cell Biology (Ministry of Education), Department of Biochemistry and Molecular Biology, China Medical University, Shenyang, 110001, China
      2. Department of Laboratory Medicine, First Affiliated Hospital, China Medical University, Shenyang, 110001, China

    • Dates

       
      Published
      December 2014
      Manuscript received
      6 April 2014
      Accepted
      22 September 2014

  • Journal of Biosciences | News

      Forthcoming Special issue.


    • To trigger further research on plant mitochondria, the Journal of Biosciences is bringing out a special issue titled "Plant Mitochondria: Properties and Interactions with Other Organelles".


      Plant mitochondria are quite distinct and have unique features, such as a cyanide-insensitive alternate pathway. They also interact with chloroplasts to optimize photosynthetic carbon assimilation.


      Submissions are welcome until 30 July 2023. The contributions can be original articles, short communications, reviews, or mini-reviews on any topic related to plant mitochondria.


      Authors can submit their articles online at https://www.editorialmanager.com/jbsc/default2.aspx

      Posted on April 12, 2023
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      Posted on July 25, 2019

      Click here for Editorial Note on CAP Mode

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