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001-es BibID:	BIBFORM004119
035-os BibID:	WOS:000253576200009
Első szerző:	Hegedűs Éva (biofizikus)
Cím:	Heteroduplex analysis using flow cytometric microbead assays to detect deletions, insertions, and single-strand lesions / Hegedüs E., Imre L., Pataki J., Lizanecz E., Székvölgyi L., Fazakas F., Bacsó Z., Tóth A., Szabó M., Seres Z., Szabó G.
Dátum:	2008
Megjegyzések:	We explore the possibilities offered by flow cytometric microbead analysis to develop high throughput methods for the detection of deletions/insertions and single-strand DNA lesions. The products of PCR reactions derived from reference and test samples are denatured and reannealed, then exposed to enzymatic or chemical treatments distinguishing homoduplices from heteroduplices. The biotin- and dye labeled reaction products are immobilized on microbeads and the homo- and heteroduplices are assessed in separate fluorescence channels, by flow cytometry. Using a model system based on the mixed lineage leukemia gene breakpoint cluster region, we demonstrate that deletions and insertions in genomic DNA can be detected, using S1 nuclease and chemical cleavage to distinguish hetero- from homoduplices, or a restriction enzyme cleaving only the homoduplices. Single-strand discontinuities can also be detected, by combining nick-translation, using labeled nucleotide, and flow cytometric microbead analysis. The methodical approaches demonstrated are applicable in a versatile manner in basic cell and molecular biological research and also promise direct application for high throughput screening of genetic diseases and lesions, including insertions or deletions of short sequence elements and single-strand lesions formed at hypersensitive sites in response to apoptotic stimuli.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban microbead heteroduplex cytometry egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A. - 73 : 3 (2008), p. 238-245. -
További szerzők:	Imre László (1979-) (biológus) Pataki Judit (1980-) (biofizikus) Lizanecz Erzsébet (1978-) (orvos) Székvölgyi Lóránt (1977-) (biofizikus, biokémikus, sejtbiológus) Fazakas Ferenc (1969-) (molekuláris biológus) Bacsó Zsolt (1963-) (biofizikus) Tóth Attila (1971-) (biológus) Szabó Miklós Seres Zoltán Szabó Gábor (1953-) (biofizikus)
Internet cím:	elektronikus változat DOI
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001-es BibID:	BIBFORM020763
035-os BibID:	(scopus)80051910015 (wos)000294938700008
Első szerző:	Imre László (biológus)
Cím:	Detection of mutations by flow cytometric melting point analysis of PCR products / Imre L., Balogh I., Kappelmayer J., Szabó M., Melegh B., Wanker E., Szabó G.
Dátum:	2011
Megjegyzések:	Exploring the possibilities offered by flow cytometric microbead analyses for the detection of genetic alterations, an assay based on the dependence of the melting point of double-stranded DNA molecules on their length has been developed, making use of PCR products carrying biotin and fluorescent moiety on their two ends. The samples of different length PCR products immobilized on streptavidine coated microbeads are heat-treated in the presence of formamide at temperatures between the melting point of the longer and that of the shorter PCR product, when the mean fluorescence intensity of the beads carrying the shorter molecules decreases as a result of denaturation, as opposed to the sample containing the longer product. The efficacy and sensitivity of the method is demonstrated in the case of the assessment of the degree of triplet expansion in Huntington's disease. Its utility for the detection of point mutations in heterozygous clinical samples is shown in the case of the BRCA1 gene. The assay is simple and may be offered for the purposes of clinical diagnostics of a number of genetic conditions. These include screening of samples for triplet expansions and SNPs predisposing for particular pathological or pharmacogenomic conditions. In general, the method described herein is offered for the diagnosis of any pathological condition where the length of a genomic or cDNA sequence is expected to be different from that of the normal allele
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk analysis article Biophysics diagnosis Dna FLOW Fluorescence Hungary Mutation Point Mutation Research Research Support Support Temperature
Megjelenés:	Cytometry. Part A. - 79 : 9 (2011), p. 720-726. -
További szerzők:	Balogh István (1972-) (molekuláris biológus, genetikus) Kappelmayer János (1960-) (laboratóriumi szakorvos) Szabó Miklós Melegh Béla Wanker Erich Szabó Gábor (1953-) (biofizikus)
Internet cím:	DOI Intézményi repozitóriumban (DEA) tárolt változat
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