Improved phenol emulsion DNA reassociation technique (PERT) using thermal cycling
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منابع مشابه
Improved phenol emulsion DNA reassociation technique (PERT) using thermal cycling.
Phenol emulsion reassociation technique (PERT) greatly enhances the rate of DNA hybridization in solution over typical hybridization conditions (1) and has successfully been used to accelerate competitive DNA reassociation for performing subtraction based genomic cloning (2-5). PERT is accomplished through the establishment and maintenance of a phenol emulsion, typically by mechanical vortexing...
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A phenol aqueous emulsion allows the reassociation of DNA a t temperatures from 6 to 68 "C. This phenol emulsion reassociation technique (PERT) also promotes the very rapid reassociation of DNA. E . coli and human DNAs a t a concentration of 4 pg/mL reacted a t room temperature with the P E R T reassociate many thousand times faster than under the standard conditions of 0.18 M Na+, 60 O C . Sol...
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Reassociating double-stranded DNA from single-stranded components is necessary for many molecular genetics experiments. The choice of a DNA reassociation method is dictated by the complexity of the starting material. Reassociation of simple oligomers needs only slow cooling in an aqueous environment, whereas reannealing the many single-stranded DNAs of complex genomic mixtures requires both a p...
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We study the renaturation of complementary single-stranded DNAs in a water-phenol two-phase system, with or without shaking. In very dilute solutions, each single-stranded DNA is strongly adsorbed at the interface at high salt concentrations. The adsorption of the single-stranded DNA is specific to phenol and relies on stacking and hydrogen bonding. We establish the interfacial nature of DNA re...
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Easy-to-use thermal cycling for performing rapid and small-volume DNA amplification on a single chip has attracted great interest in the area of rapid field detection of biological agents. For this purpose, as a more practical alternative to conventional continuous flow thermal cycling, liquid plug-flow thermal cycling utilizes a thermal gradient generated in a serpentine rectangular flow micro...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1995
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/23.12.2339