DNA Cycle Sequencing Kit is designed for DNA sequencing based on the Sanger Method (dideoxy chain termination method).
It provides a powerful tool to derive rapidly DNA and gene sequence information as required in a multitude of molecular biological and
The performance of the kit is based on a speciﬁcally engineered Taq polymerase showing an equal capability of incorporating ddNTPs and dNTPs. This
guarantees the generation of uniform and easy to read sequence band patterns at lowest background.
A minimal band compression of GC-rich DNA regions is achieved by optimally balanced termination mixtures containing 7-deaza-dGTP.
The reaction chemistry of the kit is optimized for automated DNA sequencers and requires ﬂuorescent- labeled primers.
Terminator A (blue cap): dNTP mix containing ddATP
Terminator C (blue cap): dNTP mix containing ddCTP
Terminator G (blue cap): dNTP mix containing ddGTP
Terminator T (blue cap): dNTP mix containing ddTTP
Cycle sequencing polymerase (red cap): 4units/µl
Cycle sequencing buffer (green cap): 10xconc.
Stopsolution (purple cap): 95 % formamide containing EDTA, bromophenol
blue, and xylene cyanolFF
PCR-grade water (white cap)
Shipping: shipped on blue ice
Storage Conditions: store at -20°C
Note: avoid multiple freeze / thaw cycles
Shelf Life: 12months
DNA cycle sequencing is a core technique in molecular biology allowing analysis of fmol-quantities DNA template. The enzymatic dideoxy chain termination method of Sanger relies on the linear
ampliﬁcation of a single-stranded template DNA using a single primer and thermostable polymerase. The synthesis of the complementary DNA strand starts at the speciﬁc priming site and ends with
the incorporation of a chain-terminating dideoxynucleotide triphosphate (ddNTP). This generates a multitude of fragments terminated within the desired length of the sequence. By using the four
different ddNTPs in four separate reaction vials, a set of extended primer strands terminated at each A, C, G, and T are obtained. When these fragments are separated on a suitable gel matrix the
sequence information can be read from the order of the bands.
The kit is optimized for cycle sequencing using ﬂuorescent-labeled primers. The required 5’-end ﬂuorescent label of the primer depends on the optical set-up of the used sequencing machine.
Primers should typically be 20-25 nucleotides in length with a content of 50-60 % G+C. They should be checked to avoid forming of internal duplexes or mispriming to other sites of the template.
Minimize the exposure of ﬂuorenscent- labeled primers to light.
produced in ISO-certificated company