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 A cad Sci USA 91: 3072-3076, 1994]. The partially duplex probes can be e mployed to capture single-stranded oligonucleotide targets and form pr imer-template complexes. Recently we showed that partially duplex prob es can prime Sanger sequencing reactions on immobilized, but non-ligat ed long single-stranded targets (similar to 500 nucleotide) [Fu et al. , Proc Natl Acad Sci, in press]. Here immobilized, non-ligated partial ly duplex probes were used to capture and sequence short single-strand ed targets. This strategy is capable of rapidly preparing large number s of samples for future mass spectrometric DNA sequencing.