Investigation of interaction between magnetic silica particles andlambda phage DNA fragment

Investigation of interaction between magnetic silica particles andlambda phage DNA fragment

Článek WoS

Nucleic acids belong to the most important molecules and therefore the understanding of theirproperties, function and behavior is crucial. Even though a range of analytical and biochemicalmethods have been developed for this purpose, one common step is essential for all of them –isolation of the nucleic acid from the from complex sample matrix. The use of magnetic parti-cles for the separation of nucleic acids has many advantages over other isolation methods. In thisstudy, an isolation procedure for extraction of DNA was optimized. Each step of the isolation pro-cess including washing, immobilization and elution was optimized and therefore the efficiency wasincreased from 1.7% to 28.7% and the total time was shortened from 75 to 30 min comparing to thepreviously described method. Quantification of the particular parameter influence was performed bysquare-wave voltammetry using hanging drop mercury electrode. Further, we compared the optimizedmethod with standard chloroform extraction and applied on isolation of DNA from Staphylococcus aureusand Escherichia coli.

Nucleic acids belong to the most important molecules and therefore the understanding of theirproperties, function and behavior is crucial. Even though a range of analytical and biochemicalmethods have been developed for this purpose, one common step is essential for all of them –isolation of the nucleic acid from the from complex sample matrix. The use of magnetic parti-cles for the separation of nucleic acids has many advantages over other isolation methods. In thisstudy, an isolation procedure for extraction of DNA was optimized. Each step of the isolation pro-cess including washing, immobilization and elution was optimized and therefore the efficiency wasincreased from 1.7% to 28.7% and the total time was shortened from 75 to 30 min comparing to thepreviously described method. Quantification of the particular parameter influence was performed bysquare-wave voltammetry using hanging drop mercury electrode. Further, we compared the optimizedmethod with standard chloroform extraction and applied on isolation of DNA from Staphylococcus aureusand Escherichia coli.

DNA isolation, Magnetic particles, Lambda phage, Polymerase chain reaction, Voltammetry

DNA isolation, Magnetic particles, Lambda phage, Polymerase chain reaction, Voltammetry

ŠMERKOVÁ, K.; DOSTÁLOVÁ, S.; VACULOVIČOVÁ, M.; KYNICKÝ, J.; TRNKOVÁ, L.; KRÁLÍK, M.; ADAM, V.; HUBÁLEK, J.; PROVAZNÍK, I.; KIZEK, R.

2014

01.12.2013

0731-7085

Journal of Pharmaceutical and Biomedical Analysis

86

1

Nizozemsko

65

72

13