Novel cyanine dye-labeled dideoxynucleoside triphosphates for DNA sequencing.
Single color cyanine dye-labeled (Cy 5.0 and Cy 5.5) dideoxynucleoside-5'-triphosphates, or 'terminators', containing different spacer lengths were synthesized and evaluated for efficacy in DNA sequencing methods using a modified thermally stable DNA polymerase. The single color cyanine dye terminators were formulated into two separate sets of sequencing mixes, one for Cy 5.0 and the other for Cy 5.5, and evaluated on different automated sequencing platforms. Each set of mixes included two pyrimidine terminators with 17-atom linkers and two purine terminators with 10-atom linkers between the dye and the nucleotide. The two sets of cyanine dye-labeled terminators chosen for this cycle sequencing study produced improved band patterns with band uniformity similar to that obtained with dye-primer sequencing methods.
Duthie
R Scott
RS
Amersham Biosciences, 800 Centennial Avenue, Piscataway, New Jersey 08855, USA.
Kalve
Inta M
IM
Samols
Sui Bi
SB
Hamilton
Scott
S
Livshin
Inna
I
Khot
Mahesh
M
Nampalli
Satyam
S
Kumar
Shiv
S
Fuller
Carl W
CW
D001483:Base Sequence; D002232:Carbocyanines / Q000737:chemistry; D015342:DNA Probes / Q000138:chemical synthesis*; D004259:DNA-Directed DNA Polymerase; D003854:Deoxyribonucleotides / Q000737:chemistry; D015224:Dideoxynucleosides / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006580:Genetic Carrier Screening; D006801:Humans; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000032:analysis; D017422:Sequence Analysis, DNA / Q000379:methods*
D002232:Carbocyanines; D015342:DNA Probes; D003854:Deoxyribonucleotides; D015224:Dideoxynucleosides; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D011506:Proteins; C085321:cyanine dye 5; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase