An efficient method of generate phosphatase insensitive 3' labelled DNA probes using Taq polymerase
نویسندگان
چکیده
منابع مشابه
DNA 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...
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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...
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We describe a rapid nonradioactive, double-stranded phage DNA sequencing method using ∆Taq DNA polymerase in cycle reaction for phage peptide-display library screening. This procedure is specific, rapid, sensitive and safe for the sequencing of large numbers of phage peptide-display colonies. In addition, thermal cycle sequencing with this chemiluminescent image detection protocol provides an i...
متن کاملHeat-mediated activation of affinity-immobilized Taq DNA polymerase.
A novel strategy for heat-mediated activation of recombinant Taq DNA polymerase is described. A serum albumin binding protein tag is used to affinity-immobilize an E. coli-expressed Taq DNA polymerase fusion protein onto a solid support coated with human serum albumin (HSA). Analysis of heat-mediated elution showed that elevated temperatures (> 70 degrees C) were required to significantly relea...
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...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1993
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/21.18.4413