A more stringent choice of primers can improve the performance of fluorescent automated DNA sequencers
De Bellis, G.; Manoni, M.; Pergolizzi, R.; Redolfi, M.E.; Lazzana, M.
Biotechniques 13(6): 892
1992
ISSN/ISBN: 0736-6205 PMID: 1476741 Document Number: 390024
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.