Home About us Contact
|
Home About us Contact | ||
|
|
||
Thermus Aquaticus DNA Polymerase (Thermu aquaticu + dna_polymerase)
Selected AbstractsA Population of Thermostable Reverse Transcriptases Evolved from Thermus aquaticus DNA Polymerase,I by Phage Display,ANGEWANDTE CHEMIE, Issue 7 2010Sophie Vichier-Guerre Dr. No abstract is available for this article. [source] Matrix-assisted laser desorption/ionization detection of polymerase chain reaction products by utilizing the 5,-3, exonuclease activity of Thermus aquaticus DNA polymerase,RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 6 2003N. R. Isola The 5,-3, exonuclease activity of DNA polymerase was utilized in the polymerase chain reaction system to generate a specific signal concomitant with amplification. These signals were detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). This method obviates the need to perform extensive DNA purification of reaction products that is often necessary for detecting larger DNA molecules by mass spectrometry. Oligonucleotides complementary to the internal region of the amplicon are degraded by the 5,-3, exonuclease activity and the degradation products are analyzed by MALDI mass spectrometry. We refer to this assay as the Exo-taq assay or probe degradation assay. This method should be amenable to automation. Copyright © 2003 John Wiley & Sons, Ltd. [source] Purification and characterization of Taq polymerase: A 9-week biochemistry laboratory project for undergraduate studentsBIOCHEMISTRY AND MOLECULAR BIOLOGY EDUCATION, Issue 1 2010Robert M. Bellin Abstract We have developed a 9-week undergraduate laboratory series focused on the purification and characterization of Thermus aquaticus DNA polymerase (Taq). Our aim was to provide undergraduate biochemistry students with a full-semester continuing project simulating a research-like experience, while having each week's procedure focus on a single learning goal. The laboratory series has been taught for the past 7 years, and survey-based assessment of the effectiveness of the laboratory series was completed during the 2006 and 2007 fall semesters. Statistical analysis of the survey results demonstrate that the laboratory series is very effective in teaching students the theory and practice of protein purification and analysis while also demonstrating positive results in more broad areas of scientific skill and knowledge. Amongst the findings, the largest reported increases in knowledge were related to students' understanding of how patent law relates to laboratory science, a topic of great importance to modern researchers that is readily discussed in relation to Taq polymerase. Overall, this laboratory series proves to be a very effective component in the curricula of undergraduate biology and chemistry majors and may be an appropriate laboratory experience for undergraduates. [source] One-step RNA pathogen detection with reverse transcriptase activity of a mutated thermostable Thermus aquaticus DNA polymeraseBIOTECHNOLOGY JOURNAL, Issue 2 2010Ramon Kranaster Abstract We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60°C). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at high temperatures is desired to weaken or melt secondary structure motifs. To demonstrate the ability of Taq M1 for RNA detection of pathogens, we performed TaqMan probe-based diagnostics of Dobrava viruses by one-step RT-PCR. We found similar detection sensitivities compared to commercially available RT-PCR systems without further optimization of reaction parameters, thus making this enzyme highly suitable for any PCR probe-based RNA detection method. [source] | ||||||||||