DNA polymerase is an enzyme which plays crucial role in replication and DNA repair. It found application in PCR (polymerase chain reaction) where catalyses process of in vitro DNA synthesis. To meet the demands posed by mod- ern diagnostic, molecular biology or genetic engineering it is necessary to improve DNA polymerases to obtain new or better features useful in these fields. So far implemented modifications in majority are based on improved reaction buffers, PCR enhancers and mutagenesis of these proteins. These modification lead to obtaining enzymes with higher thermostability, used for long or difficult amplicons (like GC-rich templates) or resistant to inhibitors from clinical and environmental samples. The aim of our study was determined the influence of native Taq DNA polymerase concentration on its prop- erties to amplification difficult amplicons, especially GCrich temples. Taq DNA polymerase was produced in E. coli cell, purified with using metal affinity chromatography and concentrated to a dozen units per microliter. Amplification difficult amplicons was determined by PCR with GC-rich templates in the gradient concentration of Taq DNA polymerase. Obtained results will be compared to commercial modified polymerases with particular reaction buffer for difficult amplicons. The present studies suggest that it is not necessary to used modified polymerase or improved reaction buffer to amplification difficult amplicons (GC-rich templates). A solution is used to exceeded amount of polymerase.
Autorzy
Informacje dodatkowe
- Kategoria
- Inne
- Typ
- supllement, wydanie specjalne, dodatek
- Język
- angielski
- Rok wydania
- 2015
