Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids
Joint Authors
Source
Issue
Vol. 2011, Issue 2011 (31 Dec. 2011), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2011-06-25
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
The abundance of mammalian 18S and 28S ribosomal RNA can decrease the detection sensitivity of bacterial or viral targets in complex host-pathogen mixtures.
A method to capture human RNA in a single step was developed and characterized to address this issue.
For this purpose, capture probes were covalently attached to magnetic microbeads using a dendrimer linker and the solid phase was tested using rat thymus RNA (mammalian components) with Escherichia coli RNA (bacterial target) as a model system.
Our results indicated that random capture probes demonstrated better performance than specific ones presumably by increasing the number of possible binding sites, and the use of a tetrame-thylammonium-chloride (TMA-Cl-) based buffer for the hybridization showed a beneficial effect in the selectivity.
The subtraction efficiency determined through real-time RT-PCR revealed capture-efficiencies comparable with commercially available enrichment kits.
The performance of the solid phase can be further fine tuned by modifying the annealing time and temperature.
American Psychological Association (APA)
Archer, Marie J.& Lin, Baochuan. 2011. Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids. BioMed Research International،Vol. 2011, no. 2011, pp.1-7.
https://search.emarefa.net/detail/BIM-990372
Modern Language Association (MLA)
Archer, Marie J.& Lin, Baochuan. Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids. BioMed Research International No. 2011 (2011), pp.1-7.
https://search.emarefa.net/detail/BIM-990372
American Medical Association (AMA)
Archer, Marie J.& Lin, Baochuan. Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids. BioMed Research International. 2011. Vol. 2011, no. 2011, pp.1-7.
https://search.emarefa.net/detail/BIM-990372
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-990372