Identification of mycobacterium sp. cells using PCR after isolation of PCR-ready DNA by magnetic microspheres
| 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 | Mycobacterium; Identification; PCR; P(HEMA-co-GMA) magnetic particles; |
| Description | The aim of this work was to optimize the conditions for DNA and RNA adsorption and desorption depending on the composition of the binding buffer. For this study, chicken erythrocyte DNA and calf liver RNA were used as model compounds. The final binding buffer composition was optimized using the following combinations: 4, 6, 8, and 12% PEG, and 1.0, 1.5, 2.0, and 2.5M NaCl. The binding buffer containing 8% PEG and 2M NaCl (final concentration) was shown to be the most appropriate combination for further studies. Separated DNA was eluted using TE buffer. The method developed was applied for isolation of PCR-ready DNA from mycobacterial cells using magnetic particles. Mycobacterial DNA from the eluate was detected by amplification of the IS900 and IS901 elements, special markers for the species Mycobacterium sp. avium and Mycobacterium sp. paratuberculosis. . |
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