منابع مشابه
Cell-free cloning using 29 DNA polymerase
We describe conditions for rolling-circle amplification (RCA) of individual DNA molecules 5–7 kb in size by >109-fold, using 29 DNA polymerase. The principal difficulty with amplification of small amounts of template by RCA using 29 DNA polymerase is ‘‘background’’ DNA synthesis that usually occurs when template is omitted, or at low template concentrations. Reducing the reaction volume while k...
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 کاملCell - free DNA: بیومارکر نوین تشخیص آزمایشگاهی
به انواع مختلف اسیدهای نوکلئیک آزاد در پلاسما شامل قطعات DNA ژنومی،DNA میتوکندریایی، DNA و mRNA های ویروسی و همچنین انواع miRNA اصطلاحا Cell–free nucleic acid گفته می شود. بخش مهمی از این اسیدهای نوکلئیک آزاد در پلاسما را Cell–free DNA (cfDNA) تشکیل می دهد. cfDNA به صورت قطعاتی با وزن مولکولی حدود KB 21-0.18 در سرم یا پلاسمای افراد سالم و افراد مبتلا به بسیاری از بیماری ها قابل تشخیص هستند. ...
متن کاملCloning and expression of T4 DNA polymerase.
The structural gene coding for bacteriophage T4 DNA polymerase (gene 43) has been cloned into inducible plasmid vectors, which provide a source for obtaining large amounts of this enzyme after induction. The T4 DNA polymerase produced in this fashion was purified by an innovative three-step procedure and was fully active.
متن کاملTemplate-free, polymerase-free DNA polymerization.
A combination of a DNA ligase and a restriction endonuclease provides a DNA polymerase activity, which might suggest a novel strategy for polymer synthesis.
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 2005
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.0508809102