title: A strategy for sequencing based on rolling-circle amplification on a microarray.
abstract: The sequencing of DNA fragments has become an important facet of the study of genomes. Parallel genomic DNA fragments displayed on a microarray play a key role in producing a template in next generation DNA sequencing. Here, we developed a technique to display parallel genomic DNA fragments based on the reaction with Bst DNA polymerase on a microarray. One of the hyperbranched rolling circle amplification (HRCA) primers was modified with acrylamide. HRCA products could be localized near the respective templates in polyacrylamide gel. DNA colonies produced by HRCA were displayed massively and in parallel on a microarray. Then the probe labeled with Cy 5 was hybridized to DNA colonies and extension reactions with Cy 5-labeled deoxyribonucleoside triphosphates (dNTPs) were carried out. The results show the signals of hybridization and extension reactions could be obtained. This provided a strategy for high-throughput sequencing.
authors.affiliation: College of Life Information Science and Instrument Engineering, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, P R. China. oga2001@163.com
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.forename: Jingjie
authors.initials: J
authors.lastname: Cui
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhou
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Chunxia
authors.initials: C
authors.lastname: Zhang
authors.forename: Keping
authors.initials: K
authors.lastname: Chen
authors.forename: Chaoqun
authors.initials: C
authors.lastname: Lian
authors.forename: Lihua
authors.initials: L
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Jinke
authors.initials: J
authors.lastname: Liang
mesh_terms: D001483:Base Sequence; D005453:Fluorescence; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*