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A Modified Method For Mitochondrial Purification And DNA Isolation From Muscles | ||
| Tikrit Journal of Pure Science | ||
| Article 1, Volume 11, Issue 1, 2006, Pages 246-248 | ||
| Author | ||
| Athraa H. Hassun | ||
| Abstract | ||
| Normally, the muscles are rich in mitochondria, but in myopathic patients, the muscles are degenerated, and low mitochondrial content is land marked. In the present modification a small piece of muscle biopsy (0.5-0.8gram) was taken twice at a time from each patient (12 patients with primary muscular dystrophy). The first biopsy was processed by the standard method, while the other was subjected to the modified method. In the latter method, the muscle biopsy was first minced and then digested with the enzyme trypsin III at a concentration of 0.35mg / gram for 10 minutes. In the standard method, the enzyme concentration was 0.25mg /gram, and the incubation time was 30 minutes. Therefore, some time saving was reached. But, the more important finding was highly enriched mitochondrial pellet, as well as, no contamination with other cellular components was observed. This modification has led to isolate a pure mtDNA, which was not contaminated with nuclear DNA | ||
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