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Determination of DNA sequences using peptide nucleic acid in combination with chitosan particles by maldi-top mass spectrometry |
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| รหัสดีโอไอ | |
| Title | Determination of DNA sequences using peptide nucleic acid in combination with chitosan particles by maldi-top mass spectrometry |
| Creator | Jittima Meebungpraw |
| Contributor | Voravee P. Hoven, Suda Kiatkamjornwong |
| Publisher | Chulalongkorn University |
| Publication Year | 2552 |
| Keyword | DNA, Nucleotide sequence, Peptides, Nucleic acids, Chitosan, Mass spectrometry, ดีเอ็นเอ, ลำดับนิวคลีโอไทด์, เปปไทด์, กรดนิวคลีอิก, ไคโตแซน, แมสสเปกโทรเมตรี |
| Abstract | Determination of DNA sequences is significantly important for many biotechnology-related applications ranging from medical, forensic, agriculture, and food science. The concept of using a new conformationally constrained pyrrolidinyl peptide nucleic acid (acpcPNA), anion-exchange captures in combination with MALDI-TOF mass spectrometry has been recently proven as a simple and effective way for DNA sequence analysis. This research has introduced chitosan quaternized chitosan particles as anion-exchange captures that may be applicable for the same purpose. The particles were prepared by either homogeneous or heterogeneous quaternization. The success of sub-micron, spherical, and positively charged quaternized chitosan particle formation was verified by FT-IR, 1H NMR, PCS, and TEM analyses. Investigation by MALDI-TOF mass spectrometry suggested that some quaternized particles in combination with acpcPNA were capable of detecting a single mismatched base out of 9-14 base DNA sequences. The success of selective detection generally required rinsing of the particles after capturing PNADNA hybrid by 20% formamide in phosphate buffer solution. Potential application of this technique for a synthetic DNA of which sequence mimics mutant K-ras DNA, a mutated region that is associated with cancer has also been demonstrated. |
| URL Website | cuir.car.chula.ac.th |
