Array Based PyrosequencingEdit
Pyrosequencing techniques, like many genetic tools, have been modifiedto accommodate the ever increasing amount of genomic/metagenomic data. 454 Life Sciences developed an array based pyrosequencing technology that is more adept at handling genomic/metagenomic sized data sets (1). The tehcnology utilizes magnetic beads housed in wells to bind short (400-500 bp) DNA templates. Droplets containing enzymes and deoxyribonucleotide triphosphates are applied to the beads in sequential order, and correct base pair matches will result in the emission of light as in the earlier versions of pyrosequencing. Beads can then be washed thoroughly, removing excess dNTPs, without stripping the template due to the high affinity caused by the magnetism. By utilizing multiple well arrays with large bead sets, sequencing efficiency is greatly increased. In one ten hour run 400 MB of sequence data can be generated (3. Array based pyrosequencing offers a lost cost option to scientists working with large data sets.
3. Droplet-Based Pyrosequencing Using Digital Microfluidics. Deborah J. Boles, Jonathan L. Benton, Germaine J. Siew, Miriam H. Levy, Prasanna K. Thwar, Melissa A. Sandahl, Jeremy L. Rouse, Lisa C. Perkins, Arjun P. Sudarsan, Roxana Jalili, Vamsee K. Pamula, Vijay Srinivasan, Richard B. Fair, Peter B. Griffin, Allen E. Eckhardt, and Michael G. Pollak. Analytical Chemistry 2011 83 (22), 8439-8447