Saturday, October 29, 2011
Hall 1-2 (San Jose Convention Center)
In the present discipline of marine microbiology, samples of filtered seawater from cruise expeditions must be taken back to shore for processing before any information can be obtained. The development of the Monterey Bay Aquarium Research Institute’s (MBARI) Environmental Sample Processor (ESP) is meant to accomplish molecular techniques such as DNA/RNA extractions and quantitative PCR at sea, allowing for the acquisition of real-time data. To help expand this system, we are developing primer sets for a gene expressed during iron-limiting conditions (idiA). Primers for idiA will be optimized for the genus Synechococcus, the dominant photosynthetic bacteria in the Monterey Bay site. We propose that these primer sets will differentiate between strains of Synechococcus by utilizing sequence changes in the idiA gene, allowing us to characterize the diversity in Monterey Bay.
We will use degenerate primer sets to amplify the idiA gene, using molecular cloning and sequence analysis to determine gene diversity. We will optimize primers using PCR.
The lab work process involves: PCR optimization, cloning, sequencing and sequence analysis.