Matrix-assisted laser desorption/ionization for sequencing single-stranded and double-stranded DNA.
The DNA sequence of a single-stranded and double-stranded template was determined. The templates were sequenced using the chain termination method and cycle sequencing method and detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The sequencing products were analyzed successfully without the laborious and expensive methods for removal of the template. Direct sequencing of the double-stranded template was achieved with minimal post-reaction purifications, which could be extremely important for mutation analysis and clinical diagnosis. A systematic study of the mechanisms and kinetics of sequencing reactions was also performed. The details of this analysis and directions for future improvements of the quality of sequencing are presented.
Taranenko
N I
NI
Health Sciences Research Division, Oak Ridge National Laboratory, Tennessee 37831-6378, USA.
Chung
C N
CN
Zhu
Y F
YF
Allman
S L
SL
Golovlev
V V
VV
Isola
N R
NR
Martin
S A
SA
Haff
L A
LA
Chen
C H
CH
D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000032:analysis*; D004259:DNA-Directed DNA Polymerase; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004277:DNA, Single-Stranded; D004247:DNA; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase