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Fungal Molecular Diagnostics


Traditional Sequencing Workflow

Slide 13

February 2011

The next slide depicts the typical workflow used for sequencing fungi in our laboratory. Using a biological safety cabinet, the organism to be identified is selected from a culture plate. Following lysis and a processing step that utilizes heat, the fungal DNA is amplified using a PCR reaction followed by a template cleaning step. Then a second PCR reaction is done to incorporate the dideoxynucleotides required for traditional Sanger sequencing. Following a second clean-up step to remove unincorporated nucleotides, the cycle sequencing is completed using a capillary electrophoresis instrument and the fungal DNA sequence is available for analysis by the laboratory staff.

Traditional Sequencing Workflow

 


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