Optimalizace izolace bakteriální DNA amplifikovatelné v PCR
| Title in English | Optimization of the method of PCR-ready DNA izolation |
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| Authors | |
| Year of publication | 2006 |
| Type | Article in Proceedings |
| Conference | X. Pracovní setkání biochemiků a molekulárních biologů |
| MU Faculty or unit | |
| Citation | |
| Field | Microbiology, virology |
| Keywords | DNA; magnetic particles; Lactobacillus; PCR |
| Description | The procedure was optimalised for the solid-phase bacterial DNA isolation using magnetic particles P(HEMA-co-EDMA) (1/1)covered by carboxyl groups (0,764 mM COOH/g). Magnetic silica particles (CPG, USA) were used as control. DNA adsorption was carried out in high PEG and NaCl concentrations. DNA isolated from pure cultures of bacteria of genus Lactobacillus was used. DNA elution was tested using different buffers (TE, TBE, TAE, water) and time durations including different intensity of sample mixing. The most suitable procedure was used for DNA isolation from bacteria of genus Lactobacillus in dairy products. Bacteria of genus Lactobacillus were identified using PCR with genus specific primers LbLMA 1-rev and R16-1 (Duberbet et al., 2002). The species Lactobacillus acidophilus was identified using PCR with species specific primers AciI and AciII (Tilsala et al. 1997). |
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