| Info. | Vol.5 - No.2 (2011.06.20) |
|---|---|
| Title | Detection of central single-nucleotide mismatches in short duplex DNAs on hyper-branched amine surfaces |
| Authors | Eung-Sam Kim1, Chang Ok Kim4, Joon Won Park1,2 & Kwan Yong Choi1,3 |
| Institutions | 1School of Interdisciplinary Bioscience and Bioengineering, National Core Research Center for Systems Bio-Dynamics 2Department of Chemistry, Division of Integrative Biosciences and Biotechnology 3Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea 4Division of LCD Panel, Samsung Electronics, Suwon 443-742, Korea Correspondence and requests for materials should be addressed to K.Y. Choi ( kchoi@postech.ac.kr) and J.W. Park ( jwpark@postech.ac.kr) |
| Abstract | High capability to distinguish single-nucleotide mismatches of genes using short oligonucleotide probes is essential in diagnostic methods for identification of point mutations and single nucleotide polymorphisms. To investigate the feasibility of using an aziridine-treated surface containing hyper-branched amine groups to discriminate single-nucleotide mismatches in a human gene, target probes for exons 5-8 of the p53 gene from liver cancer cells were hybridized with four types of surface-bound capture probes, one for perfect match and three for central single-nucleotide mismatches. The aziridine slide with high DNA-loading capacity exhibited greater ability to detect single-nucleotide mismatch than did the generic amine slide. When a T30 tether was linked to the capture probe, the mismatch discrimination capability increased when using a chemical cross-linker, but decreased when using UV irradiation for cross-linking. DNA duplexes had lower melting temperatures when the single-nucleotide mismatch was in the central region than when it was in the terminal region regardless of the type of mismatched nucleotide. Our results suggest that capture probes attached to the aziridine surface can effectively identify point mutations in a genomic sequence or and can estimate the affinity of gene-specific antisense oligonucleotide probes. |
| Keyword | p53 gene, Point mutation, Melting temperature, Aziridine, DNA microarray |
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