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EnglishThe discrimination of a fully matched, unlabeled KRAS wild-type (WT) (C-G) target sample with respectto three of the most frequent KRAS codon mutations (G12 S (C-A), G12 R (C-C), G12C (C-T)) was inves-tigated using an optimized detection strategy involving surface plasmon resonance (SPR), based onoptimized probe-surface density and ionic strength control. The changes observed in the SPR signal werealways larger for WT compared with the single-mismatch target DNA oligonucleotides, and were alignedwith the theoretical energy differences between the base pair C-G, C-T, C-A, C-C. Hybridization rates of∼106M−1s−1were detected without the introduction of high temperature and labels, usually neededin conventional hybridization methods. One hundred percent mutation discrimination of the matchedKRAS wild-type (C-G) sequence with respect to three mismatched G12C (C-T), G12 S (C-A), G12 R (C-C)target sequences was achieved.
| Titolo: | Ionic strength-controlled hybridization and stability of hybrids of KRAS DNA single-nucleotides: A surface plasmon resonance study | |
| Autori: | ||
| Data di pubblicazione: | 2017 | |
| Rivista: | ||
| Abstract: | The discrimination of a fully matched, unlabeled KRAS wild-type (WT) (C-G) target sample with respectto three of the most frequent KRAS codon mutations (G12 S (C-A), G12 R (C-C), G12C (C-T)) was inves-tigated using an optimized detection strategy involving surface plasmon resonance (SPR), based onoptimized probe-surface density and ionic strength control. The changes observed in the SPR signal werealways larger for WT compared with the single-mismatch target DNA oligonucleotides, and were alignedwith the theoretical energy differences between the base pair C-G, C-T, C-A, C-C. Hybridization rates of∼106M−1s−1were detected without the introduction of high temperature and labels, usually neededin conventional hybridization methods. One hundred percent mutation discrimination of the matchedKRAS wild-type (C-G) sequence with respect to three mismatched G12C (C-T), G12 S (C-A), G12 R (C-C)target sequences was achieved. | |
| Handle: | http://hdl.handle.net/11570/3148794 | |
| Appare nelle tipologie: | 14.a.1 Articolo su rivista |
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| File | Descrizione | Tipologia | Licenza | |
|---|---|---|---|---|
| 3148794.pdf | Ionic strength-controlled hybridization and stability of hybrids ofKRAS DNA single-nucleotides: A surface plasmon resonance study | Versione Editoriale (PDF) | Tutti i diritti riservati (All rights reserved) | Open Access Visualizza/Apri |
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