BCO-DMO BlogInstrument: Automated DNA Sequencer
General term for a laboratory instrument used for deciphering the order of bases in a strand of DNA. Sanger sequencers detect fluorescence from different dyes that are used to identify the A, C, G, and T extension reactions. Contemporary or Pyrosequencer methods are based on detecting the activity of DNA polymerase (a DNA synthesizing enzyme) with another chemoluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step.
Snips, approximately 2 cm in size, were taken from two random replicates within each genotype for Symbiodinium sequencing. Preliminary analyses indicate that all genotypes contain only A3 clade symbiodinium, but they may harbor different genotypes of the symbiont. These samples were flash frozed at -80C and stored until processing. DNA was isolated and extracted from 2 - 3 polyps worth of tissue using the Qiagen DNAeasy DNA isolation kit. Samples will be sent to Dr. Iliana Baums at Penn State University for microsatellite analysis and identification of possible variations in symbiont genotypes within coral genets.
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