Determination of the optimized conditions for coupling oligonucleotides with 16-mercaptohexadecanoic acid chemically adsorbed upon Au

Yi Te Wu, Jiunn-Der Liao, Je Inn Lin, Cheng-Chan Lu

研究成果: Article同行評審

3 引文 斯高帕斯(Scopus)

摘要

A specific 5′-modified amino group oligonucleotide (Primer 1), 15-mers in length, is selectively coupled with the carboxyl terminated 16-mercaptohexadecanoic acid (MHDA) chemically adsorbed on Au and subsequently hybridized with Antisense Primer. The amide-coupling process is of significance to create an intermediate structure for the purpose of adding Primer 1, while the hybridization reaction is relevant to various diagnostic purposes to determine the presence in nucleic acids for a target sequence. In this work, the coupling setting was particularly emphasized by varying commonly used temperatures and pH values with a definite concentration of coupling agents (i.e., 10 mM). The recombination with analogous hybridization treatment was investigated using high resolution X-ray photoelectron spectroscopy and a 75° grazing angle Fourier transform infrared spectrometer. On the basis of the spectroscopic studies, the optimized conditions for the coupling process that is also correlated with the molecular density of subsequent hybridization process on MHDA/Au have been proposed at 37°C and a pH value of 4.5. Therefore, it is pertinent to intensify the joining of short-chain DNA strands by complementary base pairing in diagnostic applications such as the identification of single nucleotide polymorphisms.

原文English
頁(從 - 到)1897-1904
頁數8
期刊Bioconjugate Chemistry
18
發行號6
DOIs
出版狀態Published - 2007 十一月 1

All Science Journal Classification (ASJC) codes

  • 化學 (全部)
  • 有機化學
  • 臨床生物化學
  • 生物化學、遺傳與分子生物學 (全部)
  • 生物化學

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