Use of an aqueous two-phase system and solid magnetic hydrophilic particles for isolationof PCR-ready DNA from dairy products
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Proceedings |
| Conference | 26th International symposium on Separation of Proteins, Peptides and Polynucleotides |
| MU Faculty or unit | |
| Citation | |
| Field | Microbiology, virology |
| Keywords | Bifidobacterium; Dairy products; Identification; PCR; P(HEMA-co-EDMA) magnetic particles; Two-phase system |
| Description | The identification of bacteria in real samples implies several steps: preparation of PCR-ready DNA and identification of bacterial strains by PCR. In some samples of dairy products the sensitivity of PCR was decreased due to the presence of PCR inhibitors. In these cases PCR-ready DNAs were isolated using an aqueous two-phase system, composed of 16 % (w/v) PEG 6000, NaCl and solid magnetic hydrophilic (P(HEMA-co-EDMA)) microspheres containing carboxyl groups. The method developed was applied for isolation of PCR-ready DNA from Bifidobacterium cells in some dairy products (different yoghurt samples). PCR was performed using PbiF1 and PbiR2 primers which enable amplification of a 914 bp long DNA fragment specific to the Bifidobacterium genus (Roy and Sirois, 2000). PCR products were detected using electrophoresis in 1.5 % agarose gel in TBE buffer. |
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