Difference between revisions of "Team:Georgia State/Results"
| Line 113: | Line 113: | ||
| − | + | <p>iGEM GSU Experience with Team Lambert’s Homogenizer versus the Commercial Bead Beater<p> | |
| + | <p>We did an Agrobacterium tumefaciens mediated transformation of pCB302-gfp-MBD plasmid into S. microadriaticum by first mildly disrupting the S. microadriaticum cells via bead beating and then creating a co-culture of the shaken algae cells and Agrobacterium tumefaciens carrying the plasmid of interest.<p> | ||
| + | <p>Following our collaboration with the Lambert students at the iGEM Presentation Bootcamp Meetup we hosted, we found that they had 3D-printed a low cost homogenizer of their own! So we decided to further collaborate by doing this transformation using a commercial bead beater and Lambert’s following this protocol. (link it, in wiki uploads)<p> | ||
| + | <p>We found that Lambert’s homogenizer actually worked better than the commercial one, at least for our purposes in this experiment. When we used the commercial bead beater, some of the sample leaked out during the shaking process. It should be noted the commercial bead beater was defected to some degree and wasn’t fully operational. It shook back and forth and side to side when the motion should only be a back and forth one, but alas still worked. Following the bead beating process, we observed the S. microadriaticum cells underneath the microscope and found that there were no live or even visible cells after the commercial bead beating process. We hypothesize that it was too aggressive and caused the cells to explode. However, after using Lambert’s homogenizer for this bead beating protocol, we found that the S. microadriaticum was still alive and motile. Check out the live footage of the cells below!<p> | ||
| + | <p>S. microadriaticum after Lambert Bead Beating<p> | ||
| + | <p>link this: https://www.youtube.com/watch?v=w93KRKNXa9s<p> | ||
| + | <p>S. microadriaticum following Commercial Bead Beating<p> | ||
| + | <p>link this:https://www.youtube.com/watch?v=KdY2gFSQlk8<p> | ||
| + | <p>We observed the cells once more about a week later and observed the same results.<p> | ||
| + | <p>In conclusion, for this experiment and for our purposes, Lambert’s bead beater proved to show greater cell viability following the bead beating process when compared to the commercial one we had in the lab, Cole-Parmer Mini bead beater, 115 VAC 60 Hz 5 in W x 7 in H x 10 in D (12.7 x 17.8 x 25.4 cm). Successful transformation of the S. microadriaticum is yet to be reported.<p> | ||
| + | |||
<!-- Goodbye Area Start --> | <!-- Goodbye Area Start --> | ||
<section class="akame-service-area"> | <section class="akame-service-area"> | ||
Revision as of 04:07, 21 October 2019
iGEM GSU Experience with Team Lambert’s Homogenizer versus the Commercial Bead Beater
We did an Agrobacterium tumefaciens mediated transformation of pCB302-gfp-MBD plasmid into S. microadriaticum by first mildly disrupting the S. microadriaticum cells via bead beating and then creating a co-culture of the shaken algae cells and Agrobacterium tumefaciens carrying the plasmid of interest.
Following our collaboration with the Lambert students at the iGEM Presentation Bootcamp Meetup we hosted, we found that they had 3D-printed a low cost homogenizer of their own! So we decided to further collaborate by doing this transformation using a commercial bead beater and Lambert’s following this protocol. (link it, in wiki uploads)
We found that Lambert’s homogenizer actually worked better than the commercial one, at least for our purposes in this experiment. When we used the commercial bead beater, some of the sample leaked out during the shaking process. It should be noted the commercial bead beater was defected to some degree and wasn’t fully operational. It shook back and forth and side to side when the motion should only be a back and forth one, but alas still worked. Following the bead beating process, we observed the S. microadriaticum cells underneath the microscope and found that there were no live or even visible cells after the commercial bead beating process. We hypothesize that it was too aggressive and caused the cells to explode. However, after using Lambert’s homogenizer for this bead beating protocol, we found that the S. microadriaticum was still alive and motile. Check out the live footage of the cells below!
S. microadriaticum after Lambert Bead Beating
link this: https://www.youtube.com/watch?v=w93KRKNXa9s
S. microadriaticum following Commercial Bead Beating
link this:https://www.youtube.com/watch?v=KdY2gFSQlk8
We observed the cells once more about a week later and observed the same results.
In conclusion, for this experiment and for our purposes, Lambert’s bead beater proved to show greater cell viability following the bead beating process when compared to the commercial one we had in the lab, Cole-Parmer Mini bead beater, 115 VAC 60 Hz 5 in W x 7 in H x 10 in D (12.7 x 17.8 x 25.4 cm). Successful transformation of the S. microadriaticum is yet to be reported.