Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support.
Approaches to direct solid phase sequencing of genomic and plasmid DNA have been developed using magnetic beads, coated with streptavidin, as solid support. The DNA is immobilized through selective incorporation of biotin into one of the strands. A single stranded template, suitable for sequencing, is obtained through strand-specific elution. Using this concept, in vitro amplified plasmid DNA and chromosomal DNA were sequenced directly from single colonies. The solid phase approach ensures that the amplification and the sequencing reactions can be performed under optimal conditions. The system was found to be suitable for sequencing using both isotope- and fluorescent-labelled primers.
Hultman
T
T
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Ståhl
S
S
Hornes
E
E
Uhlén
M
M
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase; D004768:Enterotoxins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005784:Gene Amplification; D005798:Genes, Bacterial; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D010957:Plasmids; D013211:Staphylococcus aureus / Q000235:genetics; D013698:Templates, Genetic
D004269:DNA, Bacterial; D004768:Enterotoxins; C026066:enterotoxin A, Staphylococcal; D004259:DNA-Directed DNA Polymerase