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Taq DNA polymerase (-dNTPs), with Robust buffer (5x200 units)

Product#: FNK-02-012-5

$120.29

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Taq DNA polymerase (Robust buffer)

Description

Thermus aquaticus DNA polymerase (Taq DNA polymerase) gene was expressed in E.coli in large quantities and highly purified. The enzyme has thermostable DNA polymerase activity and the MW is 94 kDa, same as that of the natural enzyme.

This enzyme is suitable for PCR reactions; capable of amplifying DNA with various primers.

Application

High-throughput PCR
Colony PCR
Incorporation of dUTP, dITP, and fluorescence-labeled nucleotides
Primer extension
Addition of a single nucleotide (adenosine) at the 3’-blunt ends for cloning into TA vector.

Products

Product ID	dNTP	Size	Link
FNK-02-012	-	200 units
FNK-02-012-5	-	5x200 units
FNK-02-002	+	200 units
FNK-02-002-5	+	5x200 units

General composition of PCR reaction mixture (total 50ul)

Taq DNA polymerase (5 units/ul)	*0.25 ul
10 x Standard Buffer (Taq)	5 ul
2.5mM (each) dNTPs	4ul
Template	<500ng
Primer 1	0.2～1.0uM (final conc.)
Primer 2	0.2～1.0uM (final conc.)
Sterile distilled water	up to 50ul

*Use of excess amount of enzyme is not recommended.

Storage Conditions

Taq DNA polymerase in 20mM Tris-HCl (pH 8.0), 100mM KCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.5% Tween20, 0.5% Igepal CA-630/
Store at -20℃

Concentration

5 units/ul, where one unit is defined as the amount of enzyme that can incorporate 10 nmols of total dNTPs into an acid-insoluble material in 30 minutes at 74℃ when activated salmon sperm DNA was used as template/primer.

Quality Assurance

Greater than 95% purity as determined by SDS-PAGE (CBB staining) (Fig.1) The absence of endonucleases and exonucleases was confirmed.

PCR Test

Good amplification result was obtained in PCR reaction using λDNA as a template up to 14 kB (Fig.2).

Reagents Supplied with Enzyme

1. 10 x Robust Buffer (Taq)

Fig.1 SDS-PAGE analysis of Taq DNA polymerase

Cautions for usage of Robust Buffer (Taq)

Robust Buffer induces maximum enzymatic activity. Therefore, cares should be taken to avoid production of undesirable smear bands in gel electrophoresis analysis by longer than optimal reaction time. We recommend about 5 to 10 seconds / kb elongation time for template up to 8 kb, and about 15 seconds / kb for up to 14 kb. We will recommend roughly the same elongation time to be set with 2-step PCR (shuttle PCR) and 3-step PCR. Extend the elongation time by short steps when amplification is not seen. The results of your experiments can be observed more rapidly by adopting 2-step PCR.

Protocols for PCR

Examples of PCR coditions for the amplification of various sizes of λDNA (Results shown in Fig.2)