Difference between revisions of "Team:CU-Boulder/Notebook"

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<figcaption style = "padding: 10px 10px 10px;">Fig.1 - Jonah and his creepy smile</figcaption>
 
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<p class = "main">Successful microscopy has confirmed the viability of EutS and EutC-eGFP in E.Coli, but the laser used to excite eGFP may also cause conformational change of azo-benzene. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance.Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance.</p>  
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<p class = "main">For the first week, the team participated in “iGEM boot camp” in which we quickly went through the procedures and equipment that we would be using throughout the summer. This entailed the procedures for PCR amplification, gel electrophoresis, cloning, and DNA sequencing. We extracted the Eut SMNLK via an agar stab provided from the Schmidt- Dannert lab and PCR amplified the Eut S gene using the biobrick primers and cloned on the high, low, and lac promoters onto the 1C3 backbone prior to the ligation of the Eut S gene onto these modified backbones. The 1C3 backbone was taken from the 2016 iGEM registry. Finally, we ligated the Eut S- high, low, and lac onto PSB1C3, the bio-brick standard backbone. </p>  
 
<p class = "main"><a href="#top">Go to top</a></p>
 
<p class = "main"><a href="#top">Go to top</a></p>
  

Revision as of 02:34, 29 August 2016

May
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8 9 10 11 12 13 14
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22 23 24 25 26 27 28
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June
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July
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August
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September
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October
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Notebook

Week 1: 5/8/16-5/14/16

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For the first week, the team participated in “iGEM boot camp” in which we quickly went through the procedures and equipment that we would be using throughout the summer. This entailed the procedures for PCR amplification, gel electrophoresis, cloning, and DNA sequencing. We extracted the Eut SMNLK via an agar stab provided from the Schmidt- Dannert lab and PCR amplified the Eut S gene using the biobrick primers and cloned on the high, low, and lac promoters onto the 1C3 backbone prior to the ligation of the Eut S gene onto these modified backbones. The 1C3 backbone was taken from the 2016 iGEM registry. Finally, we ligated the Eut S- high, low, and lac onto PSB1C3, the bio-brick standard backbone.

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Week 2: 5/15/16-5/21/16

Gel.1 - Lane 2: eGFP-Hi 1c3 Lane 3: eGFP-Hi 1c3 Lane 4: ladder Lane 5: eGFP-Lac 1c3 Lane 6: eGFP-Lac 1c3

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 3: 5/22/16-5/28/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 4: 5/29/16-6/4/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 5: 6/5/16-6/11/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 6: 6/12/16-6/18/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 7: 6/19/16-6/25/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 8: 6/26/16-7/2/16

Gel.2 - Lane 2: bad cut with E Lane 3: Nep-Lo 1k3 Lane 4: Nep-Lo 1k3 Lane 5: Ladder Lane 6: eGFP-Hi 1k3 Lane 7: eGFP-Hi 1k3

BSuccessful microscopy has confirmed the viability of EutS and EutC-eGFP in E.Coli, but the laser used to excite eGFP may also cause conformational change of azo-benzene. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance.Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance

Go to top

Week 9: 7/3/16-7/9/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 10

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 11: 7/10/16-7/16/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 12: 7/17/16-7/23/16

Gel.3 - Lane 2: eGFP-Hi 4A5 Lane 3: eGFP-Hi 4A5 Lane 4: Ladder Lane 5: eGFP-Hi 4A5 Lane 6: eGFP-Hi 4A5. Lane 3 and Lane 5 look good

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbbbbbbbbbbbbbbbbb

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Week 13: 7/24/16-7/30/16

Gel.4 - Lane 1: Ladder Lane 2: EutS- Hi 6A1 Lane 3: EutS-Hi 4A5 Lane 4: EutS-Hi 4A5 Lane 5: Ladder Lane 6: eGFP-Hi Lane 7: eGFP-Hi. eGFP is good

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 14: 7/31/16-8/6/16

Gel.5 - Lane 1: Ladder Lane 2: EutS-Hi Lane 3: EutS-Hi Lane 4: EutS-Hi Lane 5: Ladder Lane 6: EutS-Lac 6A1 Lane 7: EutS-Lac 4A5 Lane 8: EutS-Lac 4A5

Successful microscopy has confirmed the viability of EutS and EutC-eGFP in E.Coli, but the laser used to excite eGFP may also cause conformational change of azo-benzene. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance.Instead Neptune, another light activated protein that is excited at a much longer wavelength, will be used to visualize the formation and destruction of EutS microcompartments. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance. Future work will focus on the implementation of a multiconstruct system with EutS, EuC-Neptune, and a third construct that creates tRNA’s to integrate azo-benzene into the EutS at locations we expect to see significant steric hinderance.

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Week 15: 8/7/16-8/13/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 17: 8/14/16-8/20/16

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 18

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 19

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 20

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 21

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 22

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 23

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 24

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 25

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 26

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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Week 27

BBBBBBBBBBBBB BBBBB BBBBBBBBBBBBBBB BBBBBBBBBBBBBBBBB BBBBBBB BBBBB BBBBBBBB BBBBB bbbbbb bbb bbbbb bb bb bbbbbbbbbbb bb bbbbbbbb

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