Isolation of polymerase chain reaction-ready bacterial DNA from Lake Baikal sediments by carboxyl-functionalised magnetic polymer microspheres
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography A |
| MU Faculty or unit | |
| Citation | |
| Field | Microbiology, virology |
| Keywords | Magnetic microspheres; P(HEMA-co-EDMA); Lake Baikal sediments; Microbial DNA Magnetic separation; Polymerase chain reaction (PCR) |
| Description | Carboxyl group-containing magnetic nonporous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (P(HEMA-co-EDMA)) microspheres were used for the isolation of polymerase chain reaction(PCR)-ready DNA from samples of Baikal sediments. The occurence of false -negative results in PCR caused by the presence of extracelular inhibitors in DNA samples was solved using solid phase reversible DNA immobilisation. PCR-ready DNA was reversibly adsorbed to the microspheres in the presence of 8.0% PEG 6000 and 2.0 M sodium chloride concentrations. The adsorbed DNA was released from the microspheres in a low ioninic strength TE buffer. The quality of isolated DNA was checked by PCR amplification. |
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