Design and synthesis of new acid cleavable linkers for DNA sequencing by synthesis.
A new kind of acid sensitive tetrahydrofuranyl (THF) linker was synthesized and then reacted with 5-(6)-carboxytetramethylrhodaminesuccinimidyl ester (5(6)-TAMRA, SE), followed by di(N-succinimidyl) carbonate (DSC) and modified 2'-deoxyuridine triphosphate (dUTP); the final product, as a reversible terminator for DNA sequencing by synthesis (DNA SBS), was given obtained and confirmed by 1H-NMR, 31P-NMR, and HRMS with purity of up to 99%. The synthesized dye-labeled terminator incorporated into DNA strand successfully, and the fluorophore was cleaved completely under acidic conditions. The preliminary results encourage us to explore more acid-sensitive linkers for DNA SBS to increase the cleavage efficiency under weakly acidic conditions.
a Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine and Bio-ID Center , Shanghai Jiao Tong University , Shanghai , China.
Min
M
Jiang
Daonian
D
Tang
Xiaodong
X
Zhao
Qing
Q
Li
Yuan
Y
Zhuang
Xiaofei
X
Wei
Xiaowei
X
Li
Yazhi
Y
Liu
Xin-Yan
XY
Wu
Zhifeng
Z
Shao
Bing
B
Gong
Yu-Mei
YM
Shen
D003856:Deoxyuracil Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005663:Furans / Q000138:chemical synthesis; D012235:Rhodamines / Q000138:chemical synthesis* / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods*
D003856:Deoxyuracil Nucleotides; D005456:Fluorescent Dyes; D005663:Furans; D012235:Rhodamines
Triphosphate analogues; fluorescent base analogues; synthetic methodology