DNA polymerase, thermotoga neapolitana

Description:
Due to its thermo-stable nature, DNA polymerase, thermotoga neapolitana was identified⁽¹⁾ as an enzyme able to withstand the protein-denaturing conditions (high temperature) required during PCR⁽²⁾. Similar to Escherichia coli DNA polymerase I, but unlike Taq DNA polymerase, Tne DNA polymerase contains both 3′→5′ and 5′→3′ exonulease activity. Therefore it replaced the DNA polymerase from E. coli originally used in PCR ⁽³⁾. Taq's optimum temperature for activity is 75-80°C, with a half-life of 9 minutes at 97.5°C, and can replicate a 1000 base pair strand of DNA in less than 10 seconds at 72°C⁽⁴⁾.

Taq makes DNA products that have A (adenine) overhangs at their 3' ends. This may be useful in TA cloning, whereby a cloning vector (such as a plasmid) that has a T (thymine) 3' overhang is used, which complements with the A overhang of the PCR product, thus enabling ligation of the PCR product into the plasmid vector.

Applications:

• PCR (ordinary and high-throughput)

• Primer Extension

• Microarray Analysis

• Denaturing high performance liquid chromatography (DHPLC)

Source:
Thermotoga neapolitana (Tne) DNA polymerase belongs to the DNA polymerase I (Pol I) family.

Recommended Reaction Conditions:
94°C, 1min. -> (94°C, 10sec. -> 55°C, 30sec. ->72°C, 30sec.) for 25 cycles.

Recommended Storage Conditions:
 -20ºC  

References:

1. Chien A, Edgar DB, Trela JM (1976). "Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus". J. Bact. 127 (3): 1550–7. PMC 232952. PMID 8432.
2. 2.Saiki, RK; et al. (1988). "Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.". Science 239 (4839): 487–91. doi:10.1126/science.2448875. PMID 2448875.
3. Saiki, RK; et al. (1985). "Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia". Science 230 (4732): 1350– doi:10.1126/science.2999980. PMID 2999980.
4. Lawyer FC, et al. (1993). "High-level expression, purification, and enzymatic characterization of full-length Thermus aquaticus DNA polymerase ...". PCR Methods Appl. 2 (4): 275–87. PMID 8324500.