The LA Taq DNA Polymerase is a kind of Long PCR Polymerase that synergistically generates long PCR products with greater yield and fidelity than Taq DNA Polymerase alone. The fidelity of PCR with this enzyme is three times higher than with Taq DNA Polymerase. The LA Taq is optimized for the generation of very long amplicons: up to 40 kb with viral DNA and up to 15 kb with genomic DNA templates. The specially formulated LA Taq Buffer protects DNA from depurination and nicking during long thermal cycling. The PCR products generated with the LA Taq DNA Polymerase are mostly 3'-dA tailed, which can be cloned in TA vector.
Features
• Long PCR products
–– up to 40 kb with viral DNA as a template
–– up to15 kb with genomic DNA as a template
• Ideal for GC-rich templates up to 85% GC.
• Fidelity is three times higher than with Taq DNA Polymerase.
• High yields.
• Incorporates modified nucleotides.
Concentration
5 u/μl
Quality Control
Functionally tested in the generation of 40 kb amplicon with lambda DNA as a template.
Storage Buffer
The enzyme is supplied in: 20mM Tris-HCl (pH 8.0); 0.1mM EDTA; 1mM DTT; 100mM KCl; Stabilizers; 50% glycerol.
Storage
Store at -20°C.
Reaction PCR Mixture Set Up and Recommended thermal cycling conditions
The flowing is only an example that takes a 20KB human genomic DNA as the template, that only for reference, It can be adjusted according to the length and the sequence of the template and the primer.
Component Volume Final Concentration
Template DNA <1 ug as required
Forward Primer (10 μM) 1 μl 0.2-0.4 μM each
Reverse Primer (10 μM) 1 μl 0.2-0.4 μM each
10×LA Taq Buffer 5 μl 1×
2.5 mM dNTPs 4 μl 0.2 mM
LA Taq DNA polymerase 0.5μl 2.5 unit
ddH2O to final volume 50μl Not applicable
94℃ 3 min
94℃ 30 sec
55℃ 30 sec 30 cycles
72℃ 1 min
72℃ 5 min