Method and apparatus for enzymatic reactions performed on nucleic acid arrays conducted at dew point

The method for amplifying nucleic acid molecules from a template, comprises: (a) mixing single-stranded nucleic acid templates on a solid substrate such as a silicon wafer or glass slide, with a solution comprising an oligonucleotide primer that hybridizes to the templates and a DNA polymerase, wher...

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Hauptverfasser:	VAN NESS, JEFFREY, TABONE, JOHN C, MOYNIHAN, KRISTEN
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOIDCHEMISTRY CHEMISTRY COMBINATORIAL CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PERFORMING OPERATIONS PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS TESTING THEIR RELEVANT APPARATUS TRANSPORTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	The method for amplifying nucleic acid molecules from a template, comprises: (a) mixing single-stranded nucleic acid templates on a solid substrate such as a silicon wafer or glass slide, with a solution comprising an oligonucleotide primer that hybridizes to the templates and a DNA polymerase, wherein the mixing occurs in discrete areas on the substrate, and the solution remains in the discrete areas; (b) synthesizing a complementary strand to the template to form a duplex; (c) denaturing the duplex; and (d) synthesizing complementary strands to the template, thereby amplifying nucleic acid molecules; wherein mixing, synthesizing, and denaturing are conducted at dew point. The dew point may be maintained using an apparatus, comprising a container capable of being pressurized; a heating device; a means for generating pressure; and a means for generating saturated steam. The heating device, pressure generating means and steam generating means are controllable. The method may be adapted for detecting a single base alteration in a nucleic acid molecule, by incorporating a DNA ligase in the solution, and including a step to detect a ligation product. The first and second oligonucleotides will not ligate when there is a single base alteration at the junction base on the nucleic acid molecule. The method may also be adapted for performing single nucleotide extension assays by including a extension product detection step.