Pilot production of Taq and Pfu DNA polymerases | ||
| تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران | ||
| Article 12, Volume 17, Issue 1, September 2009, Pages 132-138 PDF (303.15 K) | ||
| Document Type: Research Paper | ||
| Author | ||
| hossein mirzaei nodoushan* | ||
| Abstract | ||
| Taq and Pfu DNA polymerases are two important enzymes in molecular biology studies. They are widely used in all molecular biology laboratories. Simple method of production and purification of the two enzymes is reported here to locally supply laboratories with high consumption level. Plasmids containing the Taq and Pfu genes were separately transformed to an E. coli strain. The transformed bacteria were grown on liquid 2x SOB culture medium for 4.5 hours. Bacterial cultures were induced to produce the enzymes. Extraction and purification of the enzymes was conducted as presented in this study. Bradford assay was used to determine the enzyme concentration. The activity and efficiency of Taq and Pfu enzymes produced here were compared to a commercial enzyme by amplifying a known genomic fragment of Arabidopsis thaliana ecotype Colombia. Therefore, the enzyme production method adopted in this study was efficient in producing high purity Taq and Pfu. | ||
| Keywords | ||
| Taq enzyme; Pfu enzyme; DNA polymerase; Gene transformation; Enzyme purification | ||
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