Difference between revisions of "Team:BGU Israel/Notebook"
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<h1>April</h1> | <h1>April</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | <p> | + | <p> |
| − | <button class="accordion">Week 9 | + | Our long-awaited microglia and astrocytes arrived! While our cell lines were getting used to their new home, we began cloning our crisper and gRNA plasmids for our two-component system. |
| + | <br/> | ||
| + | <br/> | ||
| + | <b>Highlights:</b> Received microglia and astrocytes cell lines, px601, H1, H2 CMV-dCas9-VPR, building plasmids with Gibson assembly. | ||
| + | </p> | ||
| + | <button class="accordion">Week 9</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/4/46/T--BGU_Israel--notebook_week9.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/4/46/T--BGU_Israel--notebook_week9.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 10 | + | <button class="accordion">Week 10</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/e/ea/T--BGU_Israel--notebook_week10.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/e/ea/T--BGU_Israel--notebook_week10.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 11 | + | <button class="accordion">Week 11</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/7/78/T--BGU_Israel--notebook_week11.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/7/78/T--BGU_Israel--notebook_week11.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 12 | + | <button class="accordion">Week 12</button> |
<div class="panel2"> | <div class="panel2"> | ||
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<h1>May</h1> | <h1>May</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | <p> | + | <p> |
| − | <button class="accordion">Week 13 | + | After a tireless journey, we successfully cloned most of our system’s plasmids. Now we began attempts to transfect these plasmids into our cell lines with different transfection reagents. Additionally, we began the promoter assay and astrocyte reactivity assay. |
| + | <br/> | ||
| + | <br/> | ||
| + | <b>Highlights:</b> Began Promoter assay, transfection attempts with reagents, ELISA for astrocyte reactivity assay. | ||
| + | </p> | ||
| + | <button class="accordion">Week 13</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/3/34/T--BGU_Israel--notebook_week13.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/3/34/T--BGU_Israel--notebook_week13.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 14 | + | <button class="accordion">Week 14</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/7/7d/T--BGU_Israel--notebook_week14.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/7/7d/T--BGU_Israel--notebook_week14.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 15 | + | <button class="accordion">Week 15</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/0/0c/T--BGU_Israel--notebook_week15.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/0/0c/T--BGU_Israel--notebook_week15.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 16 | + | <button class="accordion">Week 16</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/2/2d/T--BGU_Israel--notebook_week16.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/2/2d/T--BGU_Israel--notebook_week16.pdf" width="100%" height="500"> | ||
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<h1>June</h1> | <h1>June</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | <p>In | + | <p> |
| − | <button class="accordion">Week 17 | + | In June we had to face the music; our transfection was not working. We began exploring alternatives ways to insert our plasmids into our cells, such as electroporation, calcium phosphate transfection, and additional transfection reagents. |
| + | <br/> | ||
| + | <br/> | ||
| + | <b>Highlights:</b> Astrocyte Western blot, Electroporation of microglia. | ||
| + | </p> | ||
| + | <button class="accordion">Week 17</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/6/6e/T--BGU_Israel--notebook_week17.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/6/6e/T--BGU_Israel--notebook_week17.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 18 | + | <button class="accordion">Week 18</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/7/7c/T--BGU_Israel--notebook_week18.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/7/7c/T--BGU_Israel--notebook_week18.pdf" width="100%" height="500"> | ||
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| − | <button class="accordion">Week 19 | + | <button class="accordion">Week 19</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/2/22/T--BGU_Israel--notebook_week19.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/2/22/T--BGU_Israel--notebook_week19.pdf" width="100%" height="500"> | ||
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| − | <button class="accordion">Week 20 | + | <button class="accordion">Week 20</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/7/79/T--BGU_Israel--notebook_week20.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/7/79/T--BGU_Israel--notebook_week20.pdf" width="100%" height="500"> | ||
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<h1>July</h1> | <h1>July</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | <p>In | + | <p> |
| − | <button class="accordion">Week 21 | + | In July we preformed further attempts to transfect our cells and reactivity assays, and let’s not forget exams period. |
| + | <br/> | ||
| + | <br/> | ||
| + | <b>Highlights:</b> Electroporation of microglia, mycoplasma assay, colony PCR after the microglia electroporation, transfection attempts with reagents, astrocyte reactivity assay using western blot. | ||
| + | </p> | ||
| + | <button class="accordion">Week 21</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/4/47/T--BGU_Israel--notebook_week21.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/4/47/T--BGU_Israel--notebook_week21.pdf" width="100%" height="500"> | ||
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</div> | </div> | ||
| − | <button class="accordion">Week 22 | + | <button class="accordion">Week 22</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/d/db/T--BGU_Israel--notebook_week22.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/d/db/T--BGU_Israel--notebook_week22.pdf" width="100%" height="500"> | ||
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| − | <button class="accordion">Week 23 | + | <button class="accordion">Week 23</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/8/89/T--BGU_Israel--notebook_week23.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/8/89/T--BGU_Israel--notebook_week23.pdf" width="100%" height="500"> | ||
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| − | <button class="accordion">Week 24 | + | <button class="accordion">Week 24</button> |
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/f/f1/T--BGU_Israel--notebook_week24.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/f/f1/T--BGU_Israel--notebook_week24.pdf" width="100%" height="500"> | ||
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<h1>August</h1> | <h1>August</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | <p> | + | <p> |
| − | <button class="accordion">Week 25 | + | The summer was in full swing and with it our iGEM team. Each team member worked full force on experiments to reach a proof of concept. |
| + | <br/> | ||
| + | <br/> | ||
| + | <b>Highlights:</b> continued reactive astrocyte experiments. | ||
| + | </p> | ||
| + | <button class="accordion">Week 25</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
<object data="https://static.igem.org/mediawiki/2018/0/0b/T--BGU_Israel--notebook_week25.pdf" width="100%" height="500"> | <object data="https://static.igem.org/mediawiki/2018/0/0b/T--BGU_Israel--notebook_week25.pdf" width="100%" height="500"> | ||
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<h1>Experiments</h1> | <h1>Experiments</h1> | ||
<div class="col-sm-8 col-sm-offset-2"> | <div class="col-sm-8 col-sm-offset-2"> | ||
| − | < | + | <button class="accordion">Calcium Phosphate Transfection</button> |
| − | <button class="accordion"> | + | <div class="panel2"> |
| + | <object data="https://static.igem.org/mediawiki/2018/3/34/T--BGU_Israel--notebook_exp_calcium_phosphate.pdf" width="100%" height="500"> | ||
| + | <embed src="https://static.igem.org/mediawiki/2018/3/34/T--BGU_Israel--notebook_exp_calcium_phosphate.pdf" type="application/pdf" width="100%" height="600" /> | ||
| + | </object> | ||
| + | </div> | ||
| + | |||
| + | <button class="accordion">Cytokine Inhibition</button> | ||
<div class="panel2"> | <div class="panel2"> | ||
| − | <object data="https://static.igem.org/mediawiki/2018/ | + | <object data="https://static.igem.org/mediawiki/2018/d/da/T--BGU_Israel--notebook_exp_qpcr.pdf" width="100%" height="500"> |
| − | <embed src="https://static.igem.org/mediawiki/2018/ | + | <embed src="https://static.igem.org/mediawiki/2018/d/da/T--BGU_Israel--notebook_exp_qpcr.pdf" type="application/pdf" width="100%" height="600" /> |
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| − | <button class="accordion"> | + | <button class="accordion">Cell lines Infections with Lentivirus</button> |
<div class="panel2"> | <div class="panel2"> | ||
| − | <object data="https://static.igem.org/mediawiki/2018/ | + | <object data="https://static.igem.org/mediawiki/2018/0/0e/T--BGU_Israel--notebook_exp_lenti_infection.pdf" width="100%" height="500"> |
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</div> | </div> | ||
| − | <button class="accordion"> | + | <button class="accordion">Promoter Assay</button> |
<div class="panel2"> | <div class="panel2"> | ||
| − | <object data="https://static.igem.org/mediawiki/2018/ | + | <object data="https://static.igem.org/mediawiki/2018/5/51/T--BGU_Israel--notebook_exp_promoter_assay.pdf" width="100%" height="500"> |
| − | <embed src="https://static.igem.org/mediawiki/2018/ | + | <embed src="https://static.igem.org/mediawiki/2018/5/51/T--BGU_Israel--notebook_exp_promoter_assay.pdf" type="application/pdf" width="100%" height="600" /> |
</object> | </object> | ||
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| − | <button class="accordion"> | + | <button class="accordion">Reversed-Caspase-3 Induction</button> |
<div class="panel2"> | <div class="panel2"> | ||
| − | <object data="https://static.igem.org/mediawiki/2018/ | + | <object data="https://static.igem.org/mediawiki/2018/5/50/T--BGU_Israel--notebook_exp_caspase3.pdf" width="100%" height="500"> |
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Latest revision as of 16:34, 17 October 2018
Notebook
February
We began working in our brand-new lab space! When we first arrived, the lab was nothing but two work benches. We had to collect and organize the equipment, materials, cell culture room, and work area; all according to lab safety standards. We also began cloning the plasmids for our experiments and learning how to work with cell culture by practicing on HEK293 cells.
Highlights: clone Steap4, Timp1, F4/80 into pGL4.17 and pGL3 plasmid for Promoter Assay.
March
In March, we experienced our first little success. We cloned our promoters into the pGL3 vector and were able to begin our first experiment, the Promoter Assay. Meanwhile, so far, our practice cells were alive and well.
Highlights: A decision was made to concentrate our efforts on the pGL3 plasmid expression assay. Because, then we could compare the expression among the promoters and we wouldn’t have to use two sets of luciferase kits.
April
Our long-awaited microglia and astrocytes arrived! While our cell lines were getting used to their new home, we began cloning our crisper and gRNA plasmids for our two-component system.
Highlights: Received microglia and astrocytes cell lines, px601, H1, H2 CMV-dCas9-VPR, building plasmids with Gibson assembly.
May
After a tireless journey, we successfully cloned most of our system’s plasmids. Now we began attempts to transfect these plasmids into our cell lines with different transfection reagents. Additionally, we began the promoter assay and astrocyte reactivity assay.
Highlights: Began Promoter assay, transfection attempts with reagents, ELISA for astrocyte reactivity assay.
June
In June we had to face the music; our transfection was not working. We began exploring alternatives ways to insert our plasmids into our cells, such as electroporation, calcium phosphate transfection, and additional transfection reagents.
Highlights: Astrocyte Western blot, Electroporation of microglia.
July
In July we preformed further attempts to transfect our cells and reactivity assays, and let’s not forget exams period.
Highlights: Electroporation of microglia, mycoplasma assay, colony PCR after the microglia electroporation, transfection attempts with reagents, astrocyte reactivity assay using western blot.
August
The summer was in full swing and with it our iGEM team. Each team member worked full force on experiments to reach a proof of concept.
Highlights: continued reactive astrocyte experiments.
Experiments