Automated four-color DNA sequencing using primers assembled by hexamer ligation.
A procedure based on the assembly of sequencing primers by hexamer ligation and then using them in automated DNA sequencing is described. This method is based on a four-color fluorescent terminator chemistry. Sequencing ladders were analyzed using an ABI 373 DNA sequencer (Applied Biosystems, Foster City, CA, USA). The best results were obtained for primers assembled by ligation of four to ten hexamers. The accuracy of the method was estimated to be 99.5% up to 400 nt of the read sequence, and somewhat lower at 400-600 nt.
Kaczorowski
T
T
McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison 53706, USA.
Szybalski
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W
D001483:Base Sequence; D011088:DNA Ligases / Q000378:metabolism; D017931:DNA Primers / Q000378:metabolism*; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*
D017931:DNA Primers; D011088:DNA Ligases