Isolation of PCR-ready DNA from Mycobacterial cells using magnetic particles
| Authors | |
|---|---|
| Year of publication | 2005 |
| Type | Article in Proceedings |
| Conference | 11th International Symposium on Separation Sciences ISSS 2005 |
| MU Faculty or unit | |
| Citation | |
| Field | Microbiology, virology |
| Keywords | Mycobacterium; DNA; PCR |
| Description | For this study, chicken erythrocyte DNA and calf liver RNA were used. For both types of nucleic acids, we compared the binding capacity of the following particles: non-magnetic particles based on silica, magnetic particles based on ferrite, and magnetic particles P(HEMA-co-GMA) microspheres covered with carboxylic groups. A 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 knowledge acquired was used for the development of a PCR-ready mycobacterial DNA isolation technique. Mycobacterial DNA from the eluate was detected by amplification of the dnaJ gene. The primers Multidnajf and MultidnaJr amplify a product 140 bp long. The sensitivity of the reaction is 5 . 10-3 ng/ul (5.95. 10+2 copies). |
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