The fluoride cleavable 2-(cyanoethoxy)methyl (CEM) group as reversible 3'-O-terminator for DNA sequencing-by-synthesis - synthesis, incorporation, and cleavage.
A new and promising sequencing technology called sequencing-by-synthesis (SBS) enables fast determination of DNA sequences. 2'-Deoxynucleotides containing the (2-cyanoethoxy)methyl (CEM) group at the 3'-O-position are potential reversible terminators for the SBS technology. Herein we describe the synthesis, the incorporation by several polymerases, and the cleavage of this 3'-O-blocking group using 3'-O-CEM-thymidinyl-5'-O-triphosphate 7 as an example.
Földesi
A
A
Quiatech AB, Uppsala, Sweden.
Keller
A
A
Stura
A
A
Zigmantas
S
S
Kwiatkowski
M
M
Knapp
D
D
Engels
J W
JW
D000478:Alkylation; D001483:Base Sequence; D002621:Chemistry / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry; D005456:Fluorescent Dyes / Q000494:pharmacology; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000737:chemistry; D010710:Phosphates / Q000138:chemical synthesis* / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods; D013698:Templates, Genetic
D005456:Fluorescent Dyes; D009711:Nucleotides; D010710:Phosphates; D004247:DNA