A more stringent choice of primers can improve the performance of fluorescent automated DNA sequencers.
Some primers frequently used in the double-stranded dideoxy DNA sequencing technique with radioactive markers are not suited for fluorescent detection. In fact oligonucleotides have a different annealing efficiency related to their base sequence, and this is reflected in nonequivalent results, particularly in fluorescent automated DNA sequencing. We present a method for the evaluation of primer performance in automated DNA sequencers and show its application to the search for a better set of primers for pBluescript vector.
De Bellis
G
G
Consiglio Nazionale delle Richerche, Istituto di Tecnologie Biomediche Avanzate, Milano, Italy.
Manoni
M
M
Pergolizzi
R
R
Redolfi
M E
ME
Lazzana
M
M
D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D005069:Evaluation Studies as Topic; D005821:Genetic Techniques; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D013816:Thermodynamics
D015342:DNA Probes; D004247:DNA