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A more stringent choice of primers can improve the performance of fluorescent automated DNA sequencers
G De Bellis1, M Manoni, R Pergolizzi
1Consiglio Nazionale delle Richerche, Istituto di Tecnologie Biomediche Avanzate, Milano, Italy.
Biotechniques
|December 1, 1992
Abstract:
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.