High-level Expression and Purification of DNA and DNase Free Taq DNA Polymerase
نویسندگان
چکیده
منابع مشابه
Data of expression and purification of recombinant Taq DNA polymerase
Polymerase chain reaction (PCR) technique is widely used in many experimental conditions, and Taq DNA polymerase is critical in PCR process. In this article, the Taq DNA polymerase expression plasmid is reconstructed and the protein product is obtained by rapid purification, ("Rapid purification of high-activity Taq DNA polymerase" (Pluthero, 1993 [1]), "Single-step purification of a thermostab...
متن کاملRapid purification of high-activity Taq DNA polymerase.
The method described here is derived from that of Engelke et al. (1), and uses the same cloned form of Ther7nus aquaticus (Taq) DNA polymerase to produce this enzyme in E. coli. The modified purification method described here is quite simple, however it is important to note that factors such as the bacterial strain used, induction time and protein concentration during isolation have been opfimi...
متن کاملDNA sequencing using Taq polymerase.
Three DNA polymerases, namely E.coli DNA polymerase 1 (Klenow), reverse transcriptase and T7 DNA polymerase (sequenase), are commonly used for DNA sequencing by the chain termination method of Sanger and colleagues [1). However, the secondary structure of the DNA template can impede the progress of all three polymerases. I have developed a novel procedure in which the thermostable polymerase of...
متن کاملRapid purification of recombinant Taq DNA polymerase by freezing and high temperature thawing of bacterial expression cultures.
The polymerase chain reaction (PCR) and dideoxy DNA sequencing are frequently used techniques of molecular biology which utilise DNA polymerases. The high temperatures required for PCR necessitate a thermostable enzyme for DNA amplification, and DNA polymerase derived from the thermophilic microorganism, Thermus aquaticus, is used most commonly. The high optimal polymerisation temperature of th...
متن کاملCloning and Expression of Thermus Aquaticus DNA Polymerase Gene, Using a Thermo-Inducible Expression Vector
DNA polymerase gene from Thermus aquaticus strain YT1 was amplified using VENTTM DNA po-lymerase and cloned under the control of X.PR promoter and expression was induced by a shift in tern perature. The culture was then sonicated, and after centrifugation the lysate was treated with polyethyleneimine followed by a salting-out step. Finally the protein was precipitated with ammonium sulfate and...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Asian Journal of Research in Biochemistry
سال: 2018
ISSN: 2582-0516
DOI: 10.9734/ajrb/2018/v2i4607