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ŠMERKOVÁ, K. DOSTÁLOVÁ, S. ŠKUTKOVÁ, H. VACULOVIČOVÁ, M. ADAM, V. PROVAZNÍK, I. KIZEK, R.
Original Title
Isolation of Xis Gen Fragment of lambda Phage from Agarose Gel Using Magnetic Particles for Subsequent Enzymatic DNA Sequencing
Type
journal article in Web of Science
Language
English
Original Abstract
Gel electrophoresis is one of the most important methods used in biochemistry and molecular biology. The recovery of analytes from the gel required for subsequent analysis including amplification by polymerase chain reaction (PCR) or DNA sequencing is an issue due to the gel contamination. Among the other methods used for sample recovery, the application of nanomaterials is also being investigated. In this study, the applicability of magnetic particles (1 lm) for isolation of DNA fragment from agarose gel with subsequent DNA sequencing was investigated. Electrochemical analysis and DNA sequencing was used to investigate the recovery yield. The influence of dilution of the gel prior to the purification was investigated and the linear dependence with regression coefficient R2 = 0.9972 was obtained using square wave voltammetry. Moreover, bioinformatic analysis was used for comparison of obtained sequences, and simple and easy identification of nonsystematic errors caused by both fluorescence labeling reaction and electrophoretic separation. It was found that magnetic nanoparticles based isolation markedly lowered the errors occurring during sequencing of the isolated DNA fragment from 7 to 1 %.
Keywords
Gel electrophoresis, Electrochemical analysis, DNA sequencing, DNA purification, Magnetic particles
Authors
ŠMERKOVÁ, K.; DOSTÁLOVÁ, S.; ŠKUTKOVÁ, H.; VACULOVIČOVÁ, M.; ADAM, V.; PROVAZNÍK, I.; KIZEK, R.
RIV year
2012
Released
13. 10. 2012
Publisher
Springer
ISBN
0009-5893
Periodical
CHROMATOGRAPHIA
Year of study
Number
10
State
Federal Republic of Germany
Pages from
1
Pages to
6
Pages count
BibTex
@article{BUT94713, author="Kristýna {Šmerková} and Simona {Dostálová} and Helena {Vítková} and Markéta {Vaculovičová} and Vojtěch {Adam} and Valentine {Provazník} and René {Kizek}", title="Isolation of Xis Gen Fragment of lambda Phage from Agarose Gel Using Magnetic Particles for Subsequent Enzymatic DNA Sequencing", journal="CHROMATOGRAPHIA", year="2012", volume="2012", number="10", pages="1--6", doi="10.1007/s10337-012-2326-1", issn="0009-5893" }