Polyacrylamide gel film immobilized molecular beacon array for single nucleotide mismatch detection.
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.
Wang
Yijin
Y
Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, Nanjing 210096, China.
Wang
Hong
H
Gao
Lu
L
Liu
Heping
H
Lu
Zuhong
Z
He
Nongyue
N
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
D000180:Acrylic Resins; D009711:Nucleotides; D015345:Oligonucleotide Probes; D011138:Polytetrafluoroethylene; D000099166:Polyacrylamides; C016680:polyacrylamide gels