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The method of single-nucleotide variations detection using capillary electrophoresis and molecular beacons

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Abstract

We demonstrate that single-nucleotide variations in a DNA sequence can be detected using capillary electrophoresis (CE) and molecular beacons (MBs). In this method, the region surrounding the site of a nucleotide variation was amplified in a polymerase chain reaction, then hybridize PCR products with each of MBs. The sequences of the PCR products are different at the site of 2,044 in exon of interleukin (IL)-13 which to be identified. Through denaturation, the PCR product became single strand and hybridized with the completely complementary MB. The MB-target duplexes were separated using CE and solution-based fluorescence techniques. The results show that in each reaction a fluorescent response was elicited from the molecular beacon which was perfectly complementary to the amplified DNA, but not from the other MB whose probe sequence mismatched the target sequence. The method of CE based on MBs is able to identify single-nucleotide variations in a DNA sequence and can discriminate the genotyping of the SNP between the homo- and heteroduplexes of DNA fragments.

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Authors and Affiliations

  1. Department of Clinical Laboratory, The Second Affiliated Hospital of Harbin Medical University, 150086, Harbin, China

    Jinhui Wang, Yanhong Liu, Libo Duo & Lijuan Huang

  2. Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Harbin Medical University, 150086, Harbin, China

    Wei Wang

  3. Department of Clinical Laboratory, The Fourth Affiliated Hospital of Harbin Medical University, 150001, Harbin, China

    Xiaofeng Jiang

Authors
  1. Jinhui Wang
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  2. Wei Wang
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  3. Yanhong Liu
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  4. Libo Duo
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  5. Lijuan Huang
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  6. Xiaofeng Jiang
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Correspondence to Xiaofeng Jiang.

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Wang, J., Wang, W., Liu, Y. et al. The method of single-nucleotide variations detection using capillary electrophoresis and molecular beacons. Mol Biol Rep 36, 1903–1908 (2009). https://doi.org/10.1007/s11033-008-9397-y

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  • DOI: https://doi.org/10.1007/s11033-008-9397-y

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