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Abstract for fmol® DNA Cycle Sequencing System
The fmol® DNA Cycle Sequencing System is an
alternative method for enzymatic sequence analysis that takes advantage of the
intrinsic properties of the DNA polymerase isolated from Thermus aquaticus
(Taq DNA Polymerase). Thermus aquaticus is an extremely
thermophilic microorganism; its DNA polymerase shows thermal stability to 95°C.
The fmol® DNA Cycle Sequencing System includes Promega
Sequencing Grade Taq DNA Polymerase, which is a modified form that
gives superior DNA sequencing results on double-stranded DNA templates.
Sequencing Grade Taq DNA Polymerase produces a uniform band intensity,
low background and a high degree of accuracy.
The fmol® DNA Cycle Sequencing System provides the
researcher a choice of two protocols for sequencing DNA: direct incorporation of
label and end-labeled primer. The end-labeled primer protocol, a modification of
that described by Heiner et al., uses [γ-32P]ATP, [γ-33P]ATP or
[γ-35S]ATP to label the sequencing
primer. The DNA template and labeled primer are repeatedly annealed and
enzymatically extended/terminated. The end-labeled primer protocol is the most
versatile sequencing method and is useful when working with lambda DNA, PCR
templates and any template where false priming may be a problem. This protocol
generates sequence data very close to the primer and is recommended when this is
needed. |
