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Purified from E. coli strain carrying a plasmid with the cloned gene encoding Thermus aquaticus DNA polymerase. GoPhorITTM Taq DNA polymerase catalyzes 5`-3`synthesis of DNA. The enzyme lacks the 3`-5`exonuclease activity, but possesses 5`-3` exonulcease activity. GoPhorITTM Taq DNA polymerase is ideal for all routine PCR applications.
Thermus aquaticus
5 U/µL
20 mM Tris-HCl (pH 7.9), 100 mM KCl, 0.1 mM EDTA, 0.5 mM PMSF, 1 mM DTT, 50 % Glycerol
10X reation buffer with 20mM MgCI2 | 100 mM Tris-HCl (pH8.3), 500 mM KCl, 20 mM MgCl2, 1% Tween20
As most PCR products amplified with GoPhorITTM Taq have one A added at 3`-termini, if PCR fragments are to be cloned into T/A vectors, the last extension step can be extended to up to 30 min.
-20¡ÆC
PCR, Activity, SDS-PAGE/purity, endonuclease/nickase, Specific performance test
One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTPs into an acid insoluble form in 30 min at 72¡ÆC.
Single 94 kDa band was visible in SDS-PAGE.
Nicking activity, endonuclease and exonuclease activity were not detected after the incubation of 0.5 µg ¥ë-of supercoiled pUC18 DNA, 0.5 µg ¥ë-DNA or 0.5 µg ¥ë-HindIII digest with 25 units of this enzyme for 10 hours at 72¡ÆC.
PCR cloning, RT-PCR, Multiplex PCR, SSCP, RFLP, RAPD, AFLP etc
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Fig.1.
PCR-SSCP experiment using GoPhorITTM Taq DNA polymerase with supplied 10 X buffer I . The target locus is VNTR D17S5 on human genomic DNA
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Fig.2.
PCR amplification of 3 kb Lambda DNA fragment using GoPhorITTM Taq and Supplied B with serially diluted template DNA from 0.1 ng to 10 fg. M: size marker |
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