Hybridization and immobilization of long ds-DNAs on polystyrene microspheres

被引:15
|
作者
Ghosh, D
Faure, N
Kundu, S
Rondelez, F
Chatterji, D [1 ]
机构
[1] Indian Inst Sci, Mol Biophys Unit, Bangalore 560012, Karnataka, India
[2] Inst Curie, UMR CNRS 168, Lab Physico Chim Curie, Sect Rech, F-75231 Paris 05, France
关键词
D O I
10.1021/la0341963
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Here, we develop a system where long double-stranded DNAs (ds-DNAs) are immobilized on the surface of a polystyrene (PS) microsphere. A simple synthetic strategy is adopted in order to achieve this goal in which a single DNA chain is anchored by one of its extremities to a latex (PS) microsphere. We chose hybridization as a unique method to attach long ds-DNA chains in solution with oligonucleotides grafted on modified aminated polystyrene microspheres. The DNAs chosen were of various sizes and sources: T7A1 DNA (4.48 kilobase pair (kbp)), a plasmid DNA; and DeltaDIIIT7 DNA (39.34 kbp), a mutant of bacteriophage T7 DNA. The temperature dependence of the kinetics of hybridization of T7A1 DNA (4.48 kbp) in solution with an appropriate oligonucleotide (20-mer sequence) grafted on modified aminated polystyrene microspheres yielded a value of activation energy of similar to5.3 kcal/mol, consistent with a non-diffusion-controlled mechanism.
引用
收藏
页码:5830 / 5837
页数:8
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