Methods for rapid forensic analysis of mitochondrial DNA and characterization of mitochondrial DNA heteroplasmy

A forensic method of mitochondrial DNA analysis comprising the steps of:

• providing a forensic evidence sample;
• amplifying two or more segments of mitochondrial DNA obtained from said forensic evidence sample to obtain two or more amplification products;
• determining the molecular masses of said two or more amplification products by mass spectrometry, without sequencing said two or more amplification products;
• calculating base compositions of said two or more amplification products from said molecular masses;
• and
• comparing said base compositions of said two or more amplification products with at least one database comprising a plurality of known base compositions from said two or more segments of mitochondrial DNA from a plurality of subjects, thereby reaching a forensic conclusion.

A method of characterizing heteroplasmy of two or more segments of mitochondrial DNA of a subject comprising the steps of:

• providing a sample from said subject;
• amplifying said two or more segments of mitochondrial DNA from said sample with two or more primer pairs to obtain a plurality of amplification products;
• determining molecular masses of said plurality of amplification products by mass spectrometry, without sequencing said plurality of amplification products;
• and
• determining base compositions of said plurality of amplification products thereby characterizing said heteroplasmy.