We are looking to genotype our mice. Our mice have a mutation in their mitochondrial DNA and we want to know the level of heteroplasmy i.e. % of mutation in their mitochondrial DNA. So we cannot use standard sequencing to determine that and usually pyrosequencing is used to determine this. I cannot find a company that does this.
Dear Jacqueline... I hope you can benefit from this link:
https://epigendx.com/public/d/service/pyrosequencing
Hello Jacqueline. Pyrosequencing has gone out of business several years ago, so I dont think you are going to find a provider for that. Why not try Illumina NGS? See here this paper for example, where they used Illumina sequencing to assess mitochondrial heteroplasmy. Article Assessing mitochondrial heteroplasmy using next generation s...
To detect the relative level of the mito mutation vs the reference allele, you can also use droplet digital PCR (ddPCR) with probe-targeted assays labelled with FAM/VIC to quantify the absolute copy numbers of each allele. ddPCR has been widely used to quantify the level of cell-free cancer DNA mutations in the serum/blood. If you have access to a ddPCR machine from Bio-Rad or Thermo or other providers, this could be done easily. It will cost a few hundred US $ to quantify the level of mutant mito DNA molecule. Best luck with your research.