Univerzita PardubiceFakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Magnetic beads-based DNA extraction: different techniques compatible with PCR and microfluidic systems

Autoři: Srbová Jana | Dupuy Bruno | Papadakis George | Kordas Antonis | Holubová Lucie | Gizeli Electra | Bílková Zuzana

Rok: 2014

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
cze	DNA extrakce s využitím magnetických částic: srovnání technik kompatibilních s PCR a s mikrofluidními systémy	Pro extrakci bakteriální DNA byly použity různé techniky využívající magnetické částice (s terminálními silanolovými, karboxylovými i amino- skupinami). Účinnost extrakce byla hodnocena kvalitativně i kvantitativně a následně byla vyizolovaná DNA naamplifikována pomocí PCR.	magnetické částice; extrakce DNA; silika; Salmonella
eng	Magnetic beads-based DNA extraction: different techniques compatible with PCR and microfluidic systems	In this study, magnetic particles with different terminal functional groups (silanol, carboxyl and amine) and with size between 1 and 3 µm served as a solid phase for the aforementioned SPE methods. The DNA extraction efficiency from three types of materials differing in the complexity – 19 bases long oligonucleotide with Cy3 dye or gDNA of Salmonella enterica serotype Typhimurium pre-isolated or from whole-cell lysate – was monitored and compared. Moreover, specific isolation through the hybridization between the oligonucleotides immobilized on the beads and complementary sequences of the isolated DNA was performed as well. UV/VIS measurement, agarose electrophoresis, urea-PAGE and PCR were utilized for quantitative and qualitative evaluation of eluted DNA. Extraction efficiency and elution volume were another important evaluation criteria. It was confirmed that the suitability of the chosen method and particular protocol depends on the length of DNA to be isolated. The advantages and disadvantages of all methods were discussed. In future, the on-chip extraction is going to be transferred into microscale format with integrated micro-PCR module and sensor for specific detection of amplified DNA.	magnetic beads; DNA extraction; silica; Salmonella