Difference between revisions of "Team:Georgia State/Collaborations"

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                       <li class=""><a href="#Impact" data-toggle="tab"> Environmental Impact</a></li>
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                       <li class=""><a href="#Protocol" data-toggle="tab"> Emory Protocol</a></li>
 
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                                 <p class="last">Limuli plays a vital role in the ecology of estuarine and coastal communities. Horseshoe crabs utilize autochthonous and allochthonous production from pelagic and benthic food webs(figure 2; Carmichael et al. 2004). The horseshoe crab is an integral part of many vertebrate predators diet. Benthic fish feed on horseshoe crab eggs and larvae, sharks feed on the smaller juveniles, and sea turtles feed on adults (Botton et al. 2003).  Horseshoe crabs are dietary generalists, and adult crabs are ecologically essential bivalve predators in some locations. One of the most notable predator-prey relationships that were pointed out to us while we were at the Georgia Aquarium was the migratory shorebird–horseshoe crab egg interaction. Eleven species, such as the more familiar red knot(featured to the right) and the dowitcher, rely on horseshoe crab eggs for sustenance during their migration along the Atlantic Flyway (Castro and Myers 1993). Migrating birds require an estimated 539 metric tons of eggs to full the trip to the Arctic summer range(Castro and Myers 1993). To bring this into better perspective let's  consider the aforementioned red knot species.The red knot species depends so heavily on the abundance of horseshoe crab eggs that with this species depletion this bird over the last 20 years has seen a population decline from over 100,000 to less than 15,000.  Thus, the red knot is now a species nominated for protection under the Endangered Species Act. Horseshoe crabs are therefore a critical species. The horseshoe crab links an array of prey (bivalves and polychaete worms) and predators (fish, turtles, and birds), utilizing both autochthonous and allochthonous production from pelagic and benthic food webs (figure 2; Carmichael et al. 2004).</p>
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                                 <p class="last">MA6b1. cells
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- centrifuged 2500 rpm for 5 or 10 min depending on the sample
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- LB supernatant was dumped
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- cells resuspended in 30 mL of 1500 uM phosphate buffer
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- centrifuged at 2500 rpm for 10 min
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- phosphate buffer supernatant dumped
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- cells resuspended in 32 mL of 1500 uM phosphate buffer
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- 1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point
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- tubes centrifuged at 10k rpm for 5 mins at the specified time point
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    -specified time points
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        t0= 30 mins
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        t2= 1 hr
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        t3= 2.5 hrs
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        t4= 24 hrs?
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Leave overnight in a mixer at 37 C.  
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Next day, centrifuge 96 well plate.
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Recipe for Malachite Green:
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20 uL of the supernatant of each sample put into a new 96-well
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60 uL of DI water added to each well with the sample to dilute
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20 uL of Malachite green reagent added
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Malachite Green -
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500ul Malachite Green
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125ul Ammonia
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10ul Tween
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Revision as of 20:17, 1 November 2017

"It is the long history of humankind (and animal kind, too) those who learned to collaborate and improvise most effectively have prevailed."

Charles Darwin

UVA Meet-Up

The University of Virginia iGEM team invited us along with William & Mary, University of Delaware, UNC-Asheville, and the University of Maryland to the Virginia Mid-Atlantic Meetup event. Each team gave a short presentation on their project. After every two presentations, there were break-out sessions. Most of the collaboration took place during the breakout sessions. Due to this opportunity, we were able to discuss and ask questions to the other teams about their projects. The discussions helped us think about our project on an introspective level. Through our conversations with William & Mary, we realized the importance of the rate of protein expression in a vector and its relation to our protein expression rates. Analyzing how each group presented improved our overall presentation technique. After every presentation and break out session the Community Bio Labs from Charlottesville presented and then we had one final breakout session. This break-out session was the most important because we were able to speak directly with someone who had used HCG and pregnancy tests as a detection device. The conversation made us think about the concentration levels pregnancy tests detect and how we may need to alter our detection mechanism.

Emory

Emory, this year, had a cool project that focused on an issue faced by water refinery plants. Emory found during their visit to their local WaterHub that a primary concern was to come up with an efficient way to deal with constant fluctuating orthophosphate levels. Emory, to address this problem, decided to experiment on increasing the efficiency of organisms to eat phosphate. We assisted Emory in this endeavor by working with four strains of Bacillus subtilis. Two of the strains were wild-type isolated from water donated by the Emory Waterhub, and two of the strains were commercially available Bacillus subtilis. The goal of our work was to see which strain could uptake the most phosphate. To test the bacteria we the protocol provided by Emory, see tab labeled Emory protocol. Then we modified the protocol, see modified protocol. And, finally, because we were lucky enough to visit the Georgia Aquarium a place with an abundant amount of water and a state of the art filtration system we asked the aquarium for some of their unfiltered and filtered water to see if they had some of the same phosphate problems.

MA6b1. cells - centrifuged 2500 rpm for 5 or 10 min depending on the sample - LB supernatant was dumped - cells resuspended in 30 mL of 1500 uM phosphate buffer - centrifuged at 2500 rpm for 10 min - phosphate buffer supernatant dumped - cells resuspended in 32 mL of 1500 uM phosphate buffer - 1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point - tubes centrifuged at 10k rpm for 5 mins at the specified time point -specified time points t0= 30 mins t2= 1 hr t3= 2.5 hrs t4= 24 hrs? Leave overnight in a mixer at 37 C. Next day, centrifuge 96 well plate. Recipe for Malachite Green: 20 uL of the supernatant of each sample put into a new 96-well 60 uL of DI water added to each well with the sample to dilute 20 uL of Malachite green reagent added Malachite Green - 500ul Malachite Green 125ul Ammonia 10ul Tween

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