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.timeline-biology .timeline-block-left .timeline-box, .timeline-chemistry .timeline-block-left .timeline-box{ | .timeline-biology .timeline-block-left .timeline-box, .timeline-chemistry .timeline-block-left .timeline-box{ | ||
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margin-left: 10%; | margin-left: 10%; | ||
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border: none; | border: none; | ||
} | } | ||
+ | |||
+ | .timeline h3{ | ||
+ | padding-left: 10%; | ||
+ | } | ||
+ | |||
+ | .timeline-text{ | ||
+ | padding-left: 10%; | ||
+ | } | ||
+ | |||
+ | .marker{ | ||
+ | width: 45px; | ||
+ | height: 45px; | ||
+ | margin-left: 1%; | ||
+ | } | ||
+ | |||
+ | .marker p{ | ||
+ | font-size: 80%; | ||
+ | } | ||
.beam_image_name > h5 { | .beam_image_name > h5 { | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis of (Quaterthiophene-5- ylethynyl)trimethylsilane</h3> | + | <h3>Synthesis of (Quaterthiophene-5-ylethynyl)trimethylsilane</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/1/19/T--Franconia--Beam--siliane.png"> | <img src = "https://static.igem.org/mediawiki/2017/1/19/T--Franconia--Beam--siliane.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text biggerPicture"> | <div class="timeline-text biggerPicture"> | ||
− | <h3>Synthesis of α- Bromquaterthiophene</h3> | + | <h3>Synthesis of α-Bromquaterthiophene</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/5/52/T--Franconia--Beam--azide1.png"> | <img src = "https://static.igem.org/mediawiki/2017/5/52/T--Franconia--Beam--azide1.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 2-(2- (2-azidoethoxy)ethoxy)ethanol</h3> | + | <h3>Synthesis 2-(2-(2-azidoethoxy)ethoxy)ethanol</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/b/bf/T--Franconia--Beam--2222.png"> | <img src = "https://static.igem.org/mediawiki/2017/b/bf/T--Franconia--Beam--2222.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text biggerPicture"> | <div class="timeline-text biggerPicture"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text biggerPicture"> | <div class="timeline-text biggerPicture"> | ||
− | <h3>Synthesis 3,5-bis((4- iodophenyl)diazenyl)benzoic acid</h3> | + | <h3>Synthesis 3,5-bis((4-iodophenyl)diazenyl)benzoic acid</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/b/bb/T--Franconia--Beam--bromquater.png"> | <img src = "https://static.igem.org/mediawiki/2017/b/bb/T--Franconia--Beam--bromquater.png"> | ||
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</div> | </div> | ||
<div class="timeline-content"> | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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</div> | </div> | ||
</div> | </div> | ||
+ | </div> | ||
<div id = "top-post" class="timeline-block timeline-block-right"> | <div id = "top-post" class="timeline-block timeline-block-right"> | ||
<div class="marker-right"> | <div class="marker-right"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<div class="timeline-box"> | <div class="timeline-box"> | ||
<div class="timeline-text"> | <div class="timeline-text"> | ||
− | <h3>Synthesis 4-Iodo- 1-Nitrosobenzen</h3> | + | <h3>Synthesis 4-Iodo-1-Nitrosobenzen</h3> |
<div class = "title"><p>Chemical equation:</p></div> | <div class = "title"><p>Chemical equation:</p></div> | ||
<img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | <img src = "https://static.igem.org/mediawiki/2017/a/a2/T--Franconia--Beam--nitrobenzen.png"> | ||
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<h3>Timeline</h3> | <h3>Timeline</h3> | ||
<div class="timeline hidden-timeline timeline-biology"> | <div class="timeline hidden-timeline timeline-biology"> | ||
− | + | <div id = "top-post" class="timeline-block timeline-block-left"> | |
+ | <div class="marker-left"> | ||
+ | <p>01<br>Nov</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <video controls style="max-width: 100%" poster="https://static.igem.org/mediawiki/2017/thumb/b/bb/T--Franconia--Beam-VorschauDEA.jpg/1200px-T--Franconia--Beam-VorschauDEA.jpg"> | ||
+ | <source src="https://static.igem.org/mediawiki/2017/a/a8/T--Franconia--Beam-biolabDEA.mp4" type="video/mp4" /> | ||
+ | </video> | ||
+ | <div class="read-more-content"> | ||
+ | <img style="max-width: 60%; max-height: 60%" src="https://static.igem.org/mediawiki/2017/d/d8/T--Franconia--Beam-biolab01111.jpg"> | ||
+ | <p>Hydrogel with 12% gelatine, 0.1% alginate, 0.01 % salt and 87.89% azo-dye (AY38) solved water were tested.</p> | ||
+ | <img style="max-width: 60%; max-height: 60%" src="https://static.igem.org/mediawiki/2017/3/32/T--Franconia--Beam--biolab01112.jpeg"> | ||
+ | <p>After two hours irradiation of UV-Light the Hydrogel showed a fluctuation of 4mm.</p> | ||
+ | |||
+ | </div> | ||
+ | <button class="read-more"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
<div class="marker-right"> | <div class="marker-right"> | ||
<p>30<br>Oct</p></div> | <p>30<br>Oct</p></div> | ||
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- 1 µl XhoI/HindIII-HF/BssHII<br> | - 1 µl XhoI/HindIII-HF/BssHII<br> | ||
</li> | </li> | ||
+ | <li>- 2 µl pBAD/Myc-His plasmid<br> | ||
+ | - 1 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI/XhoI/HindIII-HF<br> | ||
+ | - 1 µl XhoI/HindIII-HF/BssHII<br> | ||
+ | - 5 µl H2O<br> | ||
+ | </li> | ||
+ | </ol> | ||
+ | <div class="read-more-content"> | ||
+ | <p>All insert containing samples were incubated at 37 °C for 1 h. Samples containing plasmids were dephosphorylated with alkaline phosphatase by adding:<br> | ||
+ | - 1.5 µl 10x rapid alkaline phosphatase reaction buffer<br> | ||
+ | - 0.5 µl alkaline phosphatase<br> | ||
+ | - 3 µl H2O<br> | ||
+ | The plasmid containing samples were incubated at 37 °C for 20 min. After that ligations were started with the respective inserts and plasmids:<br> | ||
+ | - 15 µl pBAD/Myc-His plasmid<br> | ||
+ | - 55 µl Insert<br> | ||
+ | - 8 µl 10x T4 DNA Ligase Buffer<br> | ||
+ | - 1 µl T4 DNA Ligase<br> | ||
+ | - 1 µl H2O<br> | ||
+ | All samples were incubated over night at room temperature. | ||
+ | </p> | ||
+ | </div> | ||
+ | <button class="read-more"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>19<br>Sep</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div><p>PCR with all constructs 1a, 2a, 3a, 4a, 5a, and 6a as template DNA with associated primers:<br></p></div> | ||
+ | <p> - 1 µl Template DNA<br> | ||
+ | - 10 µl 5x Phusion DNA Polymerase Buffer<br> | ||
+ | - 4 µl dNTPs (2,5 mM)<br> | ||
+ | - 1.5 µl DMSO<br> | ||
+ | - 2.5 µl forward primer (100 pmol/µl)<br> | ||
+ | - 2.5 µl reverse primer (100 pmol/µl)<br> | ||
+ | - 0.5 µl Phusion DNA Polymerase<br> | ||
+ | The following PCR program was used:<br> | ||
+ | - Initial denaturation: 3 min, 98 °C<br> | ||
+ | - Denaturation: 20 s, 98 °C<br> | ||
+ | - Annealing: 20 s, 56 °C<br> | ||
+ | - Elongation: 50 s, 72 °C<br> | ||
+ | - Terminal elongation: 7 min, 72 °C<br> | ||
+ | - Idle: 4 °C | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>01<br>Sep</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>New restriction digest:<br></p></div> | ||
+ | <ol> | ||
+ | <li>- 3 µl pBAD/Myc-His plasmid<br> | ||
+ | - 4 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 31 µl H2O<br> | ||
+ | </li> | ||
+ | <li>- 7 µl Insert 4a<br> | ||
+ | - 2 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 9 µl H2O<br> | ||
+ | </li> | ||
+ | <li>- 6 µl insert 5a<br> | ||
+ | - 2 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 10 µl H2O<br> | ||
+ | </li> | ||
+ | </ol> | ||
+ | <div class="read-more-content"> | ||
+ | <p>All insert containing samples were incubated at 37 °C for 1 h. The samples containing plasmids were dephosphorylated with alkaline phosphatase by adding:<br> | ||
+ | - 4.5 µl 10x rapid alkaline phosphatase reaction buffer<br> | ||
+ | - 0,5 µl alkanline phosphatase<br> | ||
+ | The plasmid samples were then incubated at 37 °C for 20 min.<br> | ||
+ | Ligation:<br> | ||
+ | Two ligations were prepared, each with one of the inserts 4a or 5a<br> | ||
+ | - 20 µl pBAD/Myc-His plasmid<br> | ||
+ | - 20 µl Insert<br> | ||
+ | - 5 µl 10x T4 DNA Ligase Buffer<br> | ||
+ | - 1 µl T4 DNA Ligase<br> | ||
+ | - 4 µl H2O<br> | ||
+ | All samples were incubated over night at room temperature. The samples were discarded due to errors in constructs. | ||
+ | </p> | ||
+ | </div> | ||
+ | <button class="read-more"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>31<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>The plasmid minipreperation and the gel electrophoresis from 30.08 were repeated.</p></div> | ||
+ | <p> | ||
+ | The gel displayed only wrong sized ligation fragments, since they run the same distance as the plasmid control. | ||
+ | </p> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/3/35/T--Franconia--Beam-biolab3108.jpg"> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>30<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Transformed plasmids from 29.08. were isolated with PROMEGA PureYield plasmid miniprep kit and a gel electrophoresis with 1 % agarose gel was run, but the control with the undigested plasmid was forgotten. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>29<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>8 colonies were picked from each plate from 28.08. to inoculate test tubes and the plates were incubated over night at 37°C.</p></div> | ||
+ | <p><br> | ||
+ | Geobacter transfer: | ||
+ | - Bottle closed with septum cap in still sealed bag | ||
+ | - Bag punctured with sterile syringe to draw in oxygen free gas | ||
+ | - Septum of Geobacter containing test tube punctured with syringe to pull in Geobacter cells | ||
+ | - Quick transfer to closed 1 L medium bottle trough septum | ||
+ | - Incubated at 30°C for cell proliferation | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>28<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>The ligation products were transformed in E.coli DH5α MCR (for protocol see 18.07.)<br></p></div> | ||
+ | <p> | ||
+ | Geobacter sulfurreducens and Geobacter metallireducens were transferred from sealed test tubes to 1 L media bottles:<br> | ||
+ | - Open Medium bottle placed in 10 L sterile bag together with sterilized septum containing cap<br> | ||
+ | - “Aerogen 2,5 L” and “Aerocult” packs activated and added to bag<br> | ||
+ | - closed bag to be airtight<br> | ||
+ | - Let stand over night<br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>25<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>Restriction digest:</p><br></div> | ||
+ | <ol> | ||
+ | <li>- 2 µl pBAD/Myc-His plasmid | ||
+ | - 3 µl Cutsmart Buffer | ||
+ | - 1 µl NcoI | ||
+ | - 1 µl XhoI | ||
+ | - 23 µl H2O | ||
+ | </li> | ||
+ | <li>- 12 µl Insert 1a | ||
+ | - 3 µl Cutsmart Buffer | ||
+ | - 1 µl NcoI | ||
+ | - 1 µl XhoI | ||
+ | - 13 µl H2O | ||
+ | </li> | ||
+ | <li>- 2 µl pBAD/Myc-His plasmid | ||
+ | - 3 µl Cutsmart Buffer | ||
+ | - 1 µl XhoI | ||
+ | - 1 µl HindIII-HF | ||
+ | - 23 µl H2O | ||
+ | </li> | ||
+ | <li>- 19 µl Insert 2a | ||
+ | - 3 µl Cutsmart Buffer | ||
+ | - 1 µl XhoI | ||
+ | - 1 µl HindIII-HF | ||
+ | - 6 µl H2O | ||
+ | </li> | ||
+ | </ol> | ||
+ | <div class="read-more-content"> | ||
+ | <p>All insert containing samples were incubated for 1h at 37 °C. The plasmid containing samples were incubated for 45 min at 37 °C. The following was added afterwards: | ||
+ | - 0,5 µl alkaline phosphatase | ||
+ | - 3,5 µl 10x rapid buffer | ||
+ | - 1 µl H2O | ||
+ | The plasmid containing samples were then incubated for another 20 min at 37°C and then heat inactivated for 10 min at 80 °C. Hereafter ligation attempts were prepared: | ||
+ | </p> | ||
+ | <ol> | ||
+ | <li>- 35 µl pBAD/Myc-His plasmid (cut with NcoI and XhoI) | ||
+ | - 30 µl Insert 1a | ||
+ | - 7 µl 10x T4 DNA Ligase Buffer | ||
+ | - 1 µl T4 DNA Ligase | ||
+ | </li> | ||
+ | <li>- 35 µl pBAD/Myc-His plasmid (cut with XhoI and HindIII-HF) | ||
+ | - 30 µl Insert 2a | ||
+ | - 7 µl 10x T4 DNA Ligase Buffer | ||
+ | - 1 µl T4 DNA Ligase | ||
+ | </li> | ||
+ | <p>All samples were incubated over night at room temperature.</p> | ||
+ | </ol> | ||
+ | </div> | ||
+ | <button class="read-more"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>18<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Gel electrophoresis with undigested Inserts and plasmid. Gel run failed</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>17<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Repeated restriction digest from 21.07. but following gel electrophoresis showed only bands of the control (see 08.08).</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>15<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>Restriction digest of isolated transformed plasmids:</p></div> | ||
+ | <p> | ||
+ | - 10 µl transformed plasmid<br> | ||
+ | - 2 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 6 µl H2O<br> | ||
+ | All samples were incubated for 1h at 37 °C. A gel electrophoresis with 1 % agarose gel was run with 6 µl 6x purple loading dye (NEB) added to each sample. Gel run failed. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>14<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>The plasmids were isolated with PROMEGA PureYield plasmid miniprep kit and stored at -20 °C.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>11<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Four colonies from each sample were picked to inoculate a test tube with 4 ml of LB-medium. The samples were incubated over night at 37 °C.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>10<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>The ligated samples were transformed (for protocol see 18.07.).</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>08<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>New cloning attempt like on 21.07. with added control sample containing:</p><br></div> | ||
+ | <p> | ||
+ | - 10 µl pBAD/Myc-His plasmid<br> | ||
+ | - 2 µl 10x T4 DNA ligase buffer<br> | ||
+ | - 1 µl T4 DNA ligase<br> | ||
+ | - 7 µl H2O<br> | ||
+ | Medium for Geobacter growth was finished. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>07<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Further weighing in of salts for Geobacter medium.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>04<br>Aug</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Further weighing in of salts for Geobacter medium.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>01<br>Aug</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Gel electrophoresis was run with digested samples from 31.07. to check enzyme activity. Gel run failed. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>31<br>Jul</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>New restriction digest (for protocol see 21.07.)</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>28<br>Jul</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Weighing in of salts for medium for Geobacter sulfurreducens (for contents see DSMZ.de -> Geobacter sulfurreducens).</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>27<br>Jul</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Repetition of the gel electrophoresis of the previous day but no change whatsoever. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>26<br>Jul</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>The transformed plasmids from the 8 tubes were isolated with PROMEGA PureYield plasmid miniprep kit. </p></div> | ||
+ | <p> | ||
+ | All samples were analysed via gel electrophoresis with 1 % agarose gel (stained with Midori green) for 45 min at 150 V. The gel displayed the bands of the transformed plasmids with inserts of 1a) and 2a) being at the same height as the pBAD/Myc-His plasmid. The transformation therefore did not work, since the expected bands needed to be higher in the gel due to their bigger size. </p> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/b/bd/T--Franconia--Beam-biolab2607.jpg"> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>25<br>Jul</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>For each sample of the plated transformation from 25.07., four colonies were partly transferred on ¼ of a new ampicillin LB-plate. | ||
+ | The rest of each colony was inoculated in a test tubes containing 4 ml of LB-medium. | ||
+ | Both the test tubes and the plates were incubated over night at 37 °C. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>24<br>Jul</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Transformation of E. coli DH5α MCR with entire samples from ligation (for protocol see 18.07.).</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>21<br>Jul</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>Restriction digest:</p><br></div> | ||
+ | <ol> | ||
+ | <li>- 10 µl pBAD/Myc-His (~ 600 ng)<br> | ||
+ | - 3 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 15 µl H2O<br> | ||
+ | </li> | ||
+ | <li>- 10 µl Insert 1a (100 ng)<br> | ||
+ | - 2 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 6 µl H2O<br> | ||
+ | </li> | ||
+ | <li>- 10 µl Insert 2a (100 ng)<br> | ||
+ | - 2 µl Cutsmart Buffer<br> | ||
+ | - 1 µl NcoI<br> | ||
+ | - 1 µl XhoI<br> | ||
+ | - 6 µl H2O<br> | ||
+ | </li> | ||
+ | </ol> | ||
+ | <div class="read-more-content"> | ||
+ | <p>All three samples were incubated at 37 °C for 1.5 h. After 1 h of incubation the pBAD/Myc-His plasmid containing sample was dephosphorylated with alkaline phosphatase by adding:<br> | ||
+ | - 3.5 µl 10x rapid alkaline phosphatase reaction buffer<br> | ||
+ | - 0.5 µl alkaline phosphatase<br> | ||
+ | - 1 µl H2O<br> | ||
+ | The sample was further incubated at 37 °C for 30 min. After an overall incubation time of 1.5 h, all contained enzymes were inactivated at 65 °C for 20 min followed by 2 min at 74 °C.<br> | ||
+ | |||
+ | Ligation for each Insert 1a and 2a:<br> | ||
+ | - 5 µl pBAD/Myc-His plasmid (100 ng)<br> | ||
+ | - 20 µl Insert (100 ng)<br> | ||
+ | - 4 µl 10x T4 DNA Ligase Buffer<br> | ||
+ | - 1 µl T4 DNA Ligase<br> | ||
+ | - 10 µl H2O<br> | ||
+ | All samples were incubated over night at room temperature.<br></p> | ||
+ | </div> | ||
+ | <button class="read-more"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>20<br>Jul</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>pBAD/Myc-His plasmids were isolated with PROMEGA PureYield plasmid miniprep kit and 15 Eppendorf tubes with ~ 65 ng/µl plasmid were obtained.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-right"> | ||
+ | <div class="marker-right"> | ||
+ | <p>19<br>Jul</p></div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <p>Three Erlenmeyer flasks (250 ml) were inoculated with each one colony from the agar plates and were shaken over night at 37°C. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="timeline-block timeline-block-left"> | ||
+ | <div class="marker-left"> | ||
+ | <p>18<br>Jul</p> | ||
+ | </div> | ||
+ | <div class="timeline-content"> | ||
+ | <div class="timeline-box"> | ||
+ | <div class="timeline-text"> | ||
+ | <div class="title"><p>Transformation of E. coli DH5α MCR with pBAD/Myc-His plasmid:<br></p></div><p> | ||
+ | - 1 µl pBAD/Myc-His plasmid was added to 100 µl of frozen competent cells<br> | ||
+ | - Incubation: on ice for 1 h<br> | ||
+ | - Heat shock: 42 °C for 1 min<br> | ||
+ | - 700 µl of preheated (37 °C) LB-medium was added<br> | ||
+ | - Incubation: 37 °C for 1 h<br> | ||
+ | - Centrifugation: 13000 x g for 2 min<br> | ||
+ | - Liquid phase was discarded<br> | ||
+ | - Cells were resuspended in remaining liquid (~ 100 µl)<br> | ||
+ | - Entire samples were plated out on ampicillin containing LB-agar plates<br> | ||
+ | - Incubation: over night at 37 °C<br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="show-timeline-wrapper"> | ||
+ | |||
+ | <div class="show-timeline">Show Timeline</div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </section> | ||
+ | |||
+ | <section class ="cd-section" id="molecularmachines"> | ||
+ | |||
+ | <div class="middle-banner molecularmachines"> | ||
+ | <div class="banner-heading"> | ||
+ | <h1>Molecular Machines</h1> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="content"> | ||
+ | <p> | ||
+ | The term “molecular machine” refers to a system that is able to perform mechanical movement on a nanoscopic scale by application of an external stimulus. In order to do so, such systems only consist of a small number of molecules. Depending on the nature of the used molecules, possible stimuli can be electrical energy (redox changes), electromagnetic energy (light) or chemical energy (change of pH value or addition of specific ions). | ||
+ | Being an exceptionally novel field of science, starting in the mid 1980’s and just being rewarded with the Nobel prize of chemistry in 2016, the current applications of molecular machines are still rather few. However, a multitude of possible applications are being explored by the minute. Many mechanical devices can be mimicked, including rotors, oscillators, gears, paddle wheels, turnstiles, brakes, ratchets and gyroscopes.[1] One example of a more sophisticated system is a molecular motor[2] that can perform a 360° rotation by subsequent application of light and temperature, as can be seen in Figure 1. In another research group, four similar rotary systems have been combined on a larger molecule in order to create a four-wheeled nano-car that was able to move over a surface upon irradiation.[3] | ||
+ | </p> | ||
+ | |||
+ | <div class="read-more-content normal-content"> | ||
+ | <img class="molecular_images" src="https://static.igem.org/mediawiki/2017/f/f8/T--Franconia--Beam-MolecularMachines11.png" width="100%"> | ||
+ | |||
+ | <p> | ||
+ | In this project, azo dyes are used as molecular machines. Upon irradiation, the -N=N- azo group changes from a trans configuration to a cis configuration. As a consequence, the distance between the substituents R is shortened, resulting in a muscle-like contraction. The working principle of azo dyes as molecular machines can be seen in Figure 2. | ||
+ | </p> | ||
+ | |||
+ | <img class="molecular_images" src="https://static.igem.org/mediawiki/2017/a/ad/T--Franconia--Beam-MolecularMachines2.png" width="100%"> | ||
+ | |||
+ | <p> | ||
+ | <br> | ||
+ | <b>References:</b> [1] V. Balzani, A. Credi, M. Venturi: Molecular Devices and Machines – A Journey into the Nano World, Wiley-VCH, Weinheim, 2003. | ||
+ | |||
+ | [2] N. Koumura, R. W. J. Zijlstra, R. A. van Delden, N. Harada, B. L. Feringa, Nature, 1999, 401, 152-155. | ||
+ | [3] T. Kudernac, N. Ruangsupapichat, M. Parschau, B. Maciá, N. Katsonis, S. R. Harutyunyan, K.-H. Ernst, B. L. Feringa, Nature, 2011, 479, 208-211. | ||
+ | </p> | ||
+ | |||
+ | </div> | ||
+ | <button class="read-more normal-content"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | |||
+ | </section> | ||
+ | |||
+ | <section class ="cd-section" id="dea"> | ||
+ | |||
+ | <div class="middle-banner dea"> | ||
+ | <div class="banner-heading"> | ||
+ | <h1>DEA</h1> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="content"> | ||
+ | <p> | ||
+ | DEAs, or dielectric elastomer actuators, generally consist of thin elastic films coated with compliant electrode material on two opposing phases. That makes them flexible plate capacitors that deform due to Maxwell stress when applying a voltage across the electrodes. Since the elastic material is incompressible, applying said voltage causes the reduction of the film in thickness to result in an expansion in area. Stacking and bundling of multiple DEAs allows an actuation system to be adapted to different loading scenarios.Several materials are suitable for manufacturing DEAs. Electrodes can be made e.g. of metals, polymers, graphite or carbon nano tubes. Elastic dielectric materials in use for industrial applications are Polyacryl, Polyurethane or Polysiloxane. This project however focusses on the synthesis of a catcher-tag polymer as dielectric medium while using carbon nano tubes for the electrodes. | ||
+ | </p> | ||
+ | <p class="read-more-content normal-content"> | ||
+ | Manufacturing DEAs is possible in multiple ways. Thereby dipping, spin-coating and casting techniques are widely spread. For biomaterials, cartridges are more suitable for handling and dispensing the dielectric material. For the application of electrodes onto the elastomer, sputter and spraying techniques are widely used within the industry. Another option is the selective wetting process based on water-based solutions for when the polymer material is hydrophobic. </br></br> | ||
+ | |||
+ | <b>References:</b> F. Nendel, S. Reitelshöfer: Conception of an infrastructure for the volumetric flow rate controlled supply of process gases and for the dynamic diversion of aerosol-flows for manufacturing flexible graphene electrodes. Institute for Factory Automation and Production Systems, (06/2017) | ||
+ | <br><br> | ||
+ | </p> | ||
+ | |||
+ | <button class="read-more normal-content"> | ||
+ | <i class="fa fa-angle-double-down"></i> | ||
+ | </button> | ||
+ | </div> | ||
+ | |||
+ | </section> |
Latest revision as of 23:06, 1 November 2017
Method 1
Protein Building Blocks
Protein building Blocks (without additional Streptavidin) . The single building blocks contain N- and C-terminal catcher-tag elements to form a permanent lock-key system. The polymer is build of two different building blocks as one single building block cannot be handled without instant polymerization. Mixing of the two building blocks start the polymerization via condensation reaction leading to a biopolymer tissue.
Pilis/Carbon Nano Tubes
Geobacter-conductive pili. The bacteria geobacter is a strictly anaerobic stem, which can produce electrically conductive proteins (Pili). Originally, they are used for their own energy gain however, they can also be used as conductive layer in our dielectric elastomer actuator and replace the carbon nanotubes as ecologically friendly alternative.
Carbon Nanotubes (CNTs) are tubular architectures made of pure carbon. Every single carbon atom is sp2 hybridized and arranged in a hexagonal pattern. One can distinguish between Single Walled Carbon Nanotubes (SWCNTs) and Multi Walled Carbon Nanotubes (MWCNTs). The fabrication of the tubes can either be achieved by laser ablation of graphite or with a catalyst in a carbon rich gas phase. As pure carbon material, they are light-weight and flexible and robust. Further their thermal and electric conductivity is highly remarkable.
Artificial Muscle
Dielectric Elastomer Actuators (DEAs) are made of alternating layers of elastic and conductive material e.g. CNTs forming a stacked capacitor. The top and bottom contacts are metallic electrodes. By application of a voltage the elastomer between the cathodes and anodes is compressed in z-dimension and expands in the x,y-plane. This leads to a muscle-like contraction of the whole system.
Method 2
Protein Building Blocks
Protein building Blocks (with additional Streptavidin). The single building blocks contain N- and C-terminal catcher-tag elements with a streptavidin in the center of the protein. The catcher-tag system binds a permanent lock-key system to each other. The streptavidin unit can bind to the biotin functionalized azo dye to integrate the molecules into the polymer tissue.
Azo Dye
Molecular Machines (Azo dyes) can fulfill motion on a molecular length scale. The contraction of the dye is driven by light, which changes the conformation of the -N=N- bond from trans to cis. Implemented in high number in a tissue they can lead to contraction of the whole tissue. The invention of such small motors was awarded the nobel prize in chemistry in 2016.
Artificial Muscle
Muscle tissue with molecular machines can be obtained by combining biopolymeric tissue with an integrated streptavidin moiety. The streptavidin can bind a biotin functionalized azo dye leading to a further cross-linking. Via light-irradiation the azo dye molecules change their conformation and contract the tissue. Thermal or irradiation with a longer wavelength restores the position of the azo dye and the tissue relaxes to the original state.
The work, which is and can be done by robots is continuously increasing as well as the number of steps where complex shaped matter has to be handled. For this purpose, soft robotics are essential to prevent damage from the material. Current materials for soft robotics are based on silicones and related polymeric materials. Combined with electrically conductive materials in alternating layers, they form dielectric elastomer actuators (DEAs), which serve as muscle in robot arms. Silicones and elastic polymers can be counted to rather cost-efficient materials. However, considering the production of the material a high amount of electricity and chemical effort must be applied. This can be circumvented by a production from E. coli, where the organism produces a economically friendly biopolymer with the desired properties. The biosynthesis of the polymer building blocks, safes a significant amount of resources and energy.
The fabrication of the silicones to a device is carried out under elevated temperatures, where high accuracy at a micrometer scale is a crucial factor. This accuracy must also be maintained for the device made of biopolymer, which can be realized with modern 3D printers. Further, the 3D-printer is operating under ambient conditions, which safes money in the fabrication process.
Due to the material properties, a special issue is the degradation of the biopolymer in dependence of time. A degradation of the material can mainly be circumvented by using the cell-free peptides, which prevents the consumption of the peptides by the cells. However, oxygen and mechanic stress are main issues, which have to be tackled. Oxygen can be excluded by instant packing of the material, whereas the mechanic durability of this material is still unknown. However, a comparison with a hydrogel polyacrylate based or a disposable DEA implies a lifetime of 100 to 2960 cycles.[1,2]
Biopolymer based DEA | Silicone based DEA | |
---|---|---|
Raw materials (price) | 6 € /g ink [3,4] | Av. 0,10 € /g ink[5] |
Fabrication | 3D printing | 3D aerosol jet printing @ 80°C[6] |
Solvent | Water | Isopropanol/terpineol[6] |
Weight | est. 1 g/mL (density of water) | 1.1-1.3 g/mL [7] |
Waste | Biodegradable polymer; Simple regain of conductive material | Recycling of silicone possible, regain of conductive material complex to achieve |
Timeline
Materials
Plasmids
- pBAD/Myc-His
- pSB1C3
Inserts/ constructs for pBAD/ Myc-His plasmid
- a) 3-Spytag-ELP5-HisTag (to be cut with NcoI and XhoI)
- a) Spytag-Streptavidin-ELP5-SnoopCatcher (to be cut with XhoI and HindIII-HF)
- a) HisTag-Snooptag-ELP5-Spycatcher (to be cut with XhoI and HindIII-HF)
- a) Kationisch kurz (to be cut with NcoI and XhoI)
- a) Anionisch kurz (to be cut with NcoI and XhoI)
- a) W51W54 (to be cut with HindIII-HF and BssHII)
Inserts/ constructs for pSB1C3 plasmid
- b) 3-Spytag-ELP5-HisTag (to be cut with EcoRI and PstI)
- b) Spytag-Streptavidin-ELP5-SnoopCatcher (to be cut with EcoRI and PstI)
- b) HisTag-Snooptag-ELP5-Spycatcher (to be cut with EcoRI and PstI)
- b) Kationisch kurz (to be cut with EcoRI and PstI)
- b) Anionisch kurz (to be cut with EcoRI and PstI)
- b) W51W54 (to be cut with EcoRI and PstI)
Timeline
The term “molecular machine” refers to a system that is able to perform mechanical movement on a nanoscopic scale by application of an external stimulus. In order to do so, such systems only consist of a small number of molecules. Depending on the nature of the used molecules, possible stimuli can be electrical energy (redox changes), electromagnetic energy (light) or chemical energy (change of pH value or addition of specific ions). Being an exceptionally novel field of science, starting in the mid 1980’s and just being rewarded with the Nobel prize of chemistry in 2016, the current applications of molecular machines are still rather few. However, a multitude of possible applications are being explored by the minute. Many mechanical devices can be mimicked, including rotors, oscillators, gears, paddle wheels, turnstiles, brakes, ratchets and gyroscopes.[1] One example of a more sophisticated system is a molecular motor[2] that can perform a 360° rotation by subsequent application of light and temperature, as can be seen in Figure 1. In another research group, four similar rotary systems have been combined on a larger molecule in order to create a four-wheeled nano-car that was able to move over a surface upon irradiation.[3]
In this project, azo dyes are used as molecular machines. Upon irradiation, the -N=N- azo group changes from a trans configuration to a cis configuration. As a consequence, the distance between the substituents R is shortened, resulting in a muscle-like contraction. The working principle of azo dyes as molecular machines can be seen in Figure 2.
References: [1] V. Balzani, A. Credi, M. Venturi: Molecular Devices and Machines – A Journey into the Nano World, Wiley-VCH, Weinheim, 2003.
[2] N. Koumura, R. W. J. Zijlstra, R. A. van Delden, N. Harada, B. L. Feringa, Nature, 1999, 401, 152-155.
[3] T. Kudernac, N. Ruangsupapichat, M. Parschau, B. Maciá, N. Katsonis, S. R. Harutyunyan, K.-H. Ernst, B. L. Feringa, Nature, 2011, 479, 208-211.
DEAs, or dielectric elastomer actuators, generally consist of thin elastic films coated with compliant electrode material on two opposing phases. That makes them flexible plate capacitors that deform due to Maxwell stress when applying a voltage across the electrodes. Since the elastic material is incompressible, applying said voltage causes the reduction of the film in thickness to result in an expansion in area. Stacking and bundling of multiple DEAs allows an actuation system to be adapted to different loading scenarios.Several materials are suitable for manufacturing DEAs. Electrodes can be made e.g. of metals, polymers, graphite or carbon nano tubes. Elastic dielectric materials in use for industrial applications are Polyacryl, Polyurethane or Polysiloxane. This project however focusses on the synthesis of a catcher-tag polymer as dielectric medium while using carbon nano tubes for the electrodes.
Manufacturing DEAs is possible in multiple ways. Thereby dipping, spin-coating and casting techniques are widely spread. For biomaterials, cartridges are more suitable for handling and dispensing the dielectric material. For the application of electrodes onto the elastomer, sputter and spraying techniques are widely used within the industry. Another option is the selective wetting process based on water-based solutions for when the polymer material is hydrophobic.
References: F. Nendel, S. Reitelshöfer: Conception of an infrastructure for the volumetric flow rate controlled supply of process gases and for the dynamic diversion of aerosol-flows for manufacturing flexible graphene electrodes. Institute for Factory Automation and Production Systems, (06/2017)