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An apparatus for sequentially combining microvolumes of reagents by infrasonic mixing

โœ Scribed by Merrill N. Camien; Robert C. Warner


Book ID
102987192
Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
582 KB
Volume
138
Category
Article
ISSN
0003-2697

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โœฆ Synopsis


A method employing high-speed infrasonic mixing for obtaining timed samples for following the progress of a moderately rapid chemical reaction is described. Drops of 10 to 50 pl each of two reagents are mixed to initiate the reaction, followed, after a measured time interval, by mixing with a drop of a third reagent to quench the reaction. The method was developed for measuring the rate of denaturation of covalently closed, circular DNA in NaOH at several temperatures. For this purpose the timed samples were analyzed by analytical ultracentrifugation. The apparatus was tested by determination of the rate of hydrolysis of 2,4dinitrophenyl acetate in an alkaline buffer. The important characteristics of the method are (i) it requires very small volumes of sample and reagents; (ii) the components of the reaction mixture are preequilibrated and mixed with no transfer outside the prescribed constant temperature environment; (iii) the mixing is very rapid, and (iv) satisfactorily precise measurements of relatively short time intervals (approximately 2 set minimum) between sequential mixings of the components are readily obtainable.


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