A cost-effective plasmid purification protocol suitable for fluorescent automated DNA sequencing.
A cost-effective, reliable, and reproducible method has been developed to produce good-quality, double-stranded plasmid DNA for automated sequence analysis. The method incorporates modifications to a previously described plasmid-purification protocol used in manual sequencing. The quality of the DNA produced from the present protocol is suitable for automated fluorescent sequencing. Using a dye-terminator sequencing protocol, most runs using plasmid DNA prepared using this protocol produced over 700 bases with greater than 99% base-calling accuracy.
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Molecular Biology Core Facility, Division of Biomedical Sciences, Meharry Medical College, Nashville, TN 37208, USA. Begobu36@ccvax.mmc.edu
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D000818:Animals; D001331:Automation; D003362:Cost-Benefit Analysis; D005456:Fluorescent Dyes; D010957:Plasmids / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA / Q000379:methods*; D014346:Trypanosoma brucei brucei / Q000235:genetics
D005456:Fluorescent Dyes