REVERSE SEQUENCING BY HYBRIDIZATION WITH FLUORESCENCE INCORPORATION
Sequencing by hybridization, one of the nextgeneration DNA sequencing techniques, can greatly accelerate biological and biomedical research. However, challenges remain in accuracy and throughput In the present study, we investigated a new strategy of DNA sequencing based on the hybridization of primers combined with fluorescence dye-tagged dideoxyribonucleotide triphosphates (ddNTPs). This simple strategy requires a universal panel of tiling primers containing multiple degenerate bases and some specific bases. The DNA sequencing involves many cycles of hybridization of primers with the immobilized template, extension with labeled ddNTPs, scanning the microarray, and removing the extended primers. Under optimal conditions, 15 bp of primer with 12 bp degenerate bases and 3 specific bases could improve the accuracy of sequencing; especially, a mismatch in the second position of the pecified bases was favorable. Additionally, as many as 64 cycles could be performed without substantial dephasing of the templates or reduction in fluorescent signals after optimizing the sequencing conditions, including the concentration and the removal of primer.
DNA sequencing SBH DNA chip
Li Yanqiang Pan Zhiqiang Yang Yao Xiao Pengfeng Lu Zuhong
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering,Southea Jiangsu Key Laboratory of Anesthesiology, Department of Anesthesiology, Xuzhou Medical College,Xuzho State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering,Southea
国际会议
重庆
英文
378-380
2011-10-28（万方平台首次上网日期，不代表论文的发表时间）