Deep Vent DNA polymerase

An especially intriguing pair of products obtained from marine organisms in recent years are Vent and Deep Vent DNA polymerase. These products are used in DNA research studies. Their special feature is that they are at least 10 times as efficient as other similar products in polymerase chain reactions because they can tolerate temperatures just below the boiling point of water, a characteristic that comparable research tools lack. Vent and Deep Vent DNA polymerases are obtained from the bacterium Thermococcus litoralis, which is found around deep-sea hydrothermal vents at the bottom of the ocean. 

For final assembly PCR, we recommend using AmpliTaq DNA polymerase which in our hands works better than Deep Vent DNA polymerase. 

X ThermoPol reaction buffer 100 mM KCl, 100 mM (NRiIjSO, 200 mMTris-HCl pH 8.8,20 mM MgSO, 1% Triton X-100 (supplied with Deep Vent DNA polymerase)... 

The PCR is a three-step cyclic process that repeatedly duplicates a specific DNA sequence, contained between two oligonucleotide sequences called primers (154,155). The two primers form the ends of the sequence of DNA to be amplified and are normally referred to as the forward and reverse primers. The forward primer is complementary to the sense strand of the DNA template and is extended 5 to 3 along the DNA by DNA polymerase enzyme (Fig. 27). The reverse primer is complementary to the antisense strand of the DNA template and is normally situated 200-500 base pairs downstream from the forward primer, although much longer sequences (up to 50 kbase) can now be amplified by PCR. The process employs a thermostable DNA polymerase enzyme (such as the Taq polymerase from Thermus aqualicus BM) extracted from bacteria found in hot water sources, such as thermal pools or deep-water vents. These enzymes are not destroyed by repeated incubation at 94 °C, the temperature at which all double stranded DNA denatures or melts to its two separate strands (155). 

If the amplified PCR product is not the correct size, the PCR reaction must be optimized (components of PCR and reaction conditions). Deep Vent polymerase can be used, but both Taq and Vent polymerase frequently increase the error rate. If DNA fragments are incompletely digested, add restriction enzyme (5 U) into digestion reaction solution or increase the incubation time. 

---