Until the development of the SequiTherm enzyme, other thermostable DNA polymerases did not generate sequence data comparable to the quality of conventional protocols. More accurate sequence data and much longer reads have been obtained using SequiTherm DNA Polymerase than could be obtained using other thermostable DNA polymerases, like Taq, for cycle sequencing. Both sequences were generated using internal labeling with 35 S-dATP and the manufacturer's recommended protocol. Figure 2 shows a comparison of sequences generated from a tRNA gene using either an IsoTherm or a Sequenase Sequencing Kit. Both sequences were generated using internal labeling with 35 S-dATP and the manufacturer's recommended protocol.
Arrows indicate comparable regions of the sequence.
Arrows 2-5 show sequencing artifacts in the Sequenase autoradiogram that are absent in the same sequence data obtained using IsoTherm DNA Polymerase. Researchers who have small amounts of template or who wish to directly sequence double-stranded DNA may prefer cycle sequencing with SequiTherm DNA Polymerase, while those who are comfortable with isothermal sequencing using the Sequenase enzyme or who do not wish to use a thermal cycler may opt for isothermal sequencing with IsoTherm DNA Polymerase. Sequencing Products catalog information.