Efficient preparation of short DNA sequence ladders potentially suitable for MALDI-TOF DNA sequencing.
Duplex probes with five-base single-stranded overhangs were developed for positional sequencing by hybridization [Broude et al., Proc Natl Acad Sci USA 91:3072-3076, 1994]. The partially duplex probes can be employed to capture single-stranded oligonucleotide targets and form primer-template complexes. Recently we showed that partially duplex probes can prime Sanger sequencing reactions on immobilized, but non-ligated long single-stranded targets (approximately 500 nucleotide) [Fu et al., Proc Natl Acad Sci, in press]. Here immobilized, non-ligated partially duplex probes were used to capture and sequence short single-stranded targets. This strategy is capable of rapidly preparing large numbers of samples for future mass spectrometric DNA sequencing.
Fu
D J
DJ
Sequenom, Inc., Boston, MA 02210, USA.
Broude
N E
NE
Köster
H
H
Smith
C L
CL
Cantor
C R
CR
D001483:Base Sequence; D015342:DNA Probes; D005240:Feasibility Studies; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D015342:DNA Probes