Isolation of Xis Gen Fragment of Emphasis Type="I

Authors: Kristyna Smerkova Simona Dostalova Helena Skutkova Marketa Ryvolova Vojtech Adam Ivo Provaznik Rene Kizek

Publish Date: 2012/10/13

Volume: 76, Issue: 7-8, Pages: 329-334

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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 μm 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 R 2 = 09972 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 Financial support from the following projects IGA IP 19/2012 NanoBioTECell GACR P102/11/1068 and CEITEC CZ105/1100/020068 is highly acknowledged The author MR wishes to express her thanks to project CZ107/2300/30039 for financial support

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