title: Polyacrylamide gel film immobilized molecular beacon array for single nucleotide mismatch detection.
abstract: We reported polyacrylamide gel immobilized molecular beacon array for single nucleotide mismatch detection in this paper. Molecular beacons are oligonucleotide probes fluorescing upon hybridization to their complementary DNA/RNA targets with excellent sensitivity and high selectivity. The specially designed molecular beacon for immobilization contains a 15 base loop sequence with a 5 base pair stem, a polyT (20 bases) spacer, a 5'-end amino group for immobilization, a fluorescein in the middle of the sequence as the fluorophore, and a 3'-end DABCYL as the quencher. Between the 5'-end amino group and the stem, the polyT is used to minimize disability caused by 5'-end immobilization. The molecular beacon microarray was fabricated by a pin-based spotting robot and the hybridization was investigated by confocal microscope. A real-time hybridization process at room temperature was registered every minute for 20 min after the target solution was pumped into the hybridization cell. The result indicates that a polyacrylamide film coated glass slide provides an ideal solution-like environment for molecular beacon probes. The potential applications of this kind of molecular beacon array are mutation detection, disease mechanisms, disease diagnostics, etc. in a parallel, cost saving, and label-free detection way.
authors: Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, Nanjing 210096, China.
authors: Yijin
authors: Y
authors: Wang
authors: Hong
authors: H
authors: Wang
authors: Lu
authors: L
authors: Gao
authors: Heping
authors: H
authors: Liu
authors: Zuhong
authors: Z
authors: Lu
authors: Nongyue
authors: N
authors: He
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry* / Q000378:metabolism; D020137:Base Pair Mismatch; D004252:DNA Mutational Analysis; D004305:Dose-Response Relationship, Drug; D005821:Genetic Techniques; D007700:Kinetics; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D009154:Mutation; D036103:Nanotechnology / Q000379:methods*; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009711:Nucleotides / Q000737:chemistry; D015345:Oligonucleotide Probes / Q000737:chemistry; D011138:Polytetrafluoroethylene / Q000737:chemistry; D013997:Time Factors; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D009711:Nucleotides; D015345:Oligonucleotide Probes; D011138:Polytetrafluoroethylene; D000099166:Polyacrylamides; C016680:polyacrylamide gels