MtDNA PCR/Sequencing Lab
Mitochondrial DNA as a Molecular Clock
This is an advanced lab that should follow another PCR lab. First, students isolate their cheek cell DNA using a saline mouthwash. After PCR amplification of their mitochondrial control region, students set up cycle sequencing reactions for their mitochondrial control region. The subsequent sequencing procedures are done at Applied Biosystems, Cold Spring Harbor, or other appropriate facility. After purifying the cycle sequencing reactions, students load their reactions on an automated DNA Sequencer. Once the data are generated, they are posted on the Sequence Server at the DNA Learning Center. Students download their data and analyze individual and class results at school. Analysis includes ClustalW sequence alignment between students, primates, and ancient DNA including Neandertal. Finally, students will calculate the time since human divergence.
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