ROLLING CIRCLE AMPLIFICATION DETECTION OF RNA AND DNA

Rolling circle amplification (RCA) has been useful for detecting point mutations in isolated nucleic acids, but its application in cytological preparations has been problematic. By pretreating cells with a combination of restriction enzymes and exonucleases, we demonstrate RCA in solution and in sit...

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Hauptverfasser:	CHRISTIAN, ALLEN, T, PATTEE, MELISSA, S, ATTIX, CRISTINA, M, TUCKER, JAMES, D
Format:	Patent
Sprache:	eng ; fre
Schlagworte:	BEER BIOCHEMISTRY 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 MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Zusammenfassung:	Rolling circle amplification (RCA) has been useful for detecting point mutations in isolated nucleic acids, but its application in cytological preparations has been problematic. By pretreating cells with a combination of restriction enzymes and exonucleases, we demonstrate RCA in solution and in situ to detect gene copy number and single base mutations. It can also detect and quantify transcribed RNA in individual cells, making it a versatile tool for cell-based assays. L'amplification en cercle roulant (RCA) est utile pour détecter des mutations ponctuelles dans des acides nucléiques isolés, mais son application dans des préparations cytologiques est problématique. En pré-traitant des cellules au moyen d'une association d'enzymes de restriction et d'exonucléases, on prouve l'utilité de l'amplification en cercle roulant en solution et in situ pour détecter le nombre de copies d'un gène et des mutations de monobases. Cette méthode d'amplification peut également détecter et quantifier des ARN transcrits dans des cellules individuelles, ce qui en fait un outil polyvalent, utile dans des dosages cellulaires.