Immobilization of self-quenched DNA hairpin probe with a heterobifunctional reagent on a glass surface for sensitive detection of oligonucleotides.
A new sensitive method for the detection of nucleic acids on a glass surface has been described. The self-quenched DNA hairpin probe is immobilized on a glass surface utilizing heterobifunctional reagent, N-(3-triethoxysilylpropyl)-4-(isothiocyanatomethyl)-cyclohexane-1-carboxamide (TPICC). In the closed state fluorescence intensity was quenched due to the presence of guanosine residues in close vicinity of fluorophore while on hybridization with perfectly matched complementary target strand fluorescence was restored. Efficiency and specificity of immobilization as well as thermal stability at variable temperature and pH conditions have been discussed in detail. The method employed has potential for the detection of single nucleotide variations and other diagnostic studies.
Misra
Arvind
A
Department of Chemistry, Banaras Hindu University, Varanasi, India. arvindmisra2003@yahoo.com
Shahid
Mohammad
M
D003510:Cyclohexanes / Q000138:chemical synthesis / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry* / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry; D005898:Glass / Q000737:chemistry*; D017879:Isothiocyanates / Q000138:chemical synthesis / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry
D003510:Cyclohexanes; D015342:DNA Probes; D005456:Fluorescent Dyes; D017879:Isothiocyanates; C532456:N-(3-triethoxysilylpropyl)-4-(isothiocyanatomethyl)cyclohexane-1-carboxamide; D009841:Oligonucleotides