Optimal procedure for isolation of mutant mitochondrial alleles
US6027883A · kind A · utility
7Cited by
3References
23Claims
0Family size
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Key dates
| Filing date | Nov 27, 1996 |
| Grant date | Feb 22, 2000 |
| Priority date | — |
| Expiry date | Nov 27, 2016 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/16
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A sample preparation procedure for mitochondrial (mt) DNA analysis is described. The present method for isolating mtDNA uses sedimentation techniques for separating erythrocytes from lymphocytes and platelets (crude buffy coat fraction), followed by DNA extraction from the crude buffy coat fraction by boiling in water. This approach improves the yield of mutant DNA, enhancing the sensitivity of subsequent mutation interrogation techniques and allowing for meaningful statistical treatment of the degree of heteroplasmy within the mitochondrial DNA.
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