Mitochondrial DNA from Lumbriculus variegatus: Isolation and Restriction Digest Analysis
Gary Oxford
Tested Studies in Laboratory Teaching, 2001, Volume 22
Abstract
In this activity, mitochondria are isolated from the California blackworm, Lumbriculus variegatus, by homogenization and differential centrifugation. A modified plasmid miniprep procedure (alkaline lysis) is then used to isolate DNA from these mitochondria. Single and double enzyme restriction digests of mitochondrial DNA (mtDNA) are analyzed by agarose gel electrophoresis. Sizes of mtDNA restriction fragments are estimated from gel results and are used to determine the total size of the L. variegatus mitochondrial genome. Issues involved in creating a restriction map of mtDNA from this organism are discussed.
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