Extraction of PCR-ready DNA from Staphylococcus aureus bacteriophages using carboxyl functionalized magnetic nonporous microspheres.
Magnetic microspheres P(HEMA-co-EDMA) were used for PCR-ready phage DNA isolation from lysogenic strains of Staphylococcus aureus, including two new clinical isolates. The conditions of phage particle lysis were optimized. The quality of eluted phage DNA was evaluated by PCR. It was demonstrated that PCR-ready phage DNA can be isolated from small volumes of phage lysates (150 microl) by magnetic microspheres. The reported method is very expeditious without using toxic compounds such as phenol or chloroform. It can be used for phage identification and phage gene detection.
Kahánková
Jana
J
Masaryk University, Faculty of Science, Institute of Experimental Biology, Tvrdého 14, CZ-602 00 Brno, Czech Republic.
Spanová
Alena
A
Pantůcek
Roman
R
Horák
Daniel
D
Doskar
Jirí
J
Rittich
Bohuslav
B
D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D005821:Genetic Techniques; D008280:Magnetics; D008863:Microspheres; D016133:Polymerase Chain Reaction; D052616:Solid Phase Extraction / Q000379:methods*; D013204:Staphylococcus Phages / Q000235:genetics* / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000821:virology
D004279:DNA, Viral