Optimixation of per for amplification of D1S80 (PMCT118) allelles by using vent DNA polymerase.
- Author:
Giang Hoai Nguyen
- Publication Type:Journal Article
- Keywords:
allele
- MeSH:
DNA polymerase;
Alleles
- From:Journal of Medical Research
2001;16(3):20-23
- CountryViet Nam
- Language:Vietnamese
-
Abstract:
D1S80 locus is highly polymorphic and has been used worldwide as an important marker for forensic, medical analysis and paternity tests. Vent DNA Polymerase is a high-fidelity thermophilic DNA polymerase. The fidelity of Vent DNA polymerase is 5 to 15 folds higher than that observed for Tag DNA polymerase. This high fidelity derives in part from an integral 3’-5’ proofreading exonuclease activity in Vent DNA polymerase. Greater than 90% of the polymerase activity remains after one hour incubation at 95oC. Because of these advantages, we want to apply Vent DNA polymerase for the amplification of D1S80 alleles by PCR. Our study showed that PCR condition for Vent DNA polymerase are rather different than for Tag DNA polymerase. The amplification of D1S80 alleles with Vent DNA polymerase is optimal with the following parameters: 1. A final concentration of MgSO4 in PCR mixture of 4 mM. 2. An annealing temperature for the specific D1S80 primers 68oC. 3. A final concentration of dNTPs of 200mcM.
