TĚŠÍNSKÁ, Iva, Alena ŠPANOVÁ, Bohuslav RITTICH and Milan BARTOŠ. Identification of mycobacterium sp. cells using PCR after isolation of PCR-ready DNA by magnetic microspheres. In 26th International symposium on Separation of Proteins, Peptides and Polynucleotides. Innsbruck: Dechema, 2006, p. 77-77.
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Basic information
Original name Identification of mycobacterium sp. cells using PCR after isolation of PCR-ready DNA by magnetic microspheres
Name in Czech Identifikace buněk Mycobacterium sp. pomocí PCR po izolaci DNA pomocí magnetických mikročástic
Authors TĚŠÍNSKÁ, Iva (203 Czech Republic), Alena ŠPANOVÁ (203 Czech Republic), Bohuslav RITTICH (203 Czech Republic, guarantor) and Milan BARTOŠ (203 Czech Republic).
Edition Innsbruck, 26th International symposium on Separation of Proteins, Peptides and Polynucleotides, p. 77-77, 1 pp. 2006.
Publisher Dechema
Other information
Original language English
Type of outcome Proceedings paper
Field of Study 10600 1.6 Biological sciences
Country of publisher Austria
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14310/06:00016212
Organization unit Faculty of Science
Keywords in English Mycobacterium; Identification; PCR; P(HEMA-co-GMA) magnetic particles;
Tags identification, MYCOBACTERIUM, P(HEMA-co-GMA) magnetic particles, PCR
Tags International impact
Changed by Changed by: doc. Ing. Bohuslav Rittich, CSc., učo 1639. Changed: 5/1/2007 18:42.
Abstract
The aim of this work was to optimize the conditions for DNA and RNA adsorption and desorption depending on the composition of the binding buffer. For this study, chicken erythrocyte DNA and calf liver RNA were used as model compounds. The final binding buffer composition was optimized using the following combinations: 4, 6, 8, and 12% PEG, and 1.0, 1.5, 2.0, and 2.5M NaCl. The binding buffer containing 8% PEG and 2M NaCl (final concentration) was shown to be the most appropriate combination for further studies. Separated DNA was eluted using TE buffer. The method developed was applied for isolation of PCR-ready DNA from mycobacterial cells using magnetic particles. Mycobacterial DNA from the eluate was detected by amplification of the IS900 and IS901 elements, special markers for the species Mycobacterium sp. avium and Mycobacterium sp. paratuberculosis. .
Abstract (in Czech)
Cílem této práce bylo optimalizovat pro adsorpci a desorpci v závislosti na složení vazebného pufru.DNA z kuřecích erytrocytů a RNA z hovězích jater byly použity jako modelové sloučeniny. Optimalizace vazebného pufru byla provedena z následujících kombinací: 4, 6, 8, a 12% PEG; a 1,0, 1,5, 2,0, and 2,5 M NaCl. Vazebný pufr obsahující 8 % PEG a 2 M NaCl byl shledán jako nejvhodnější kombinace pro další studue. Separovaná DNAbyla eluována pomocí TE pufru. Vyvinutý postup byl využit pro izolaci DNA v kvalitě pro PCR z buněk mykobakterií za použitímagnetických částic. DNA v eluátu byla detekována amplifikací IS900 a IS901 elementů, speciciálních markerů pro druhy Mycobacterium sp. avium a Mycobacterium sp. paratuberculosis.
Links
GA203/05/2256, research and development projectName: Magnetické hydrofilní polymerní mikročástice reagující na vnější stimuly pro lékařství a bioinženýrství
Investor: Czech Science Foundation, Magnetic stimuli-responsive hydrophilic polymer microspheres for biomedicine/bioengineering
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