Difference between revisions of "Team:IIT Delhi/Notebook"

 
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     <div class="dropdown-content">
 
       <a href="/Team:IIT_Delhi/Circuit_Design">Circuit design and construction</a>
 
       <a href="/Team:IIT_Delhi/Circuit_Design">Circuit design and construction</a>
       <a href="/Team:IIT_Delhi/Microfluidics">Microfluidics and Fluroscence</a>
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       <a href="/Team:IIT_Delhi/Microfluidics">Microfluidics and Fluorescence</a>
 
       <a href="/Team:IIT_Delhi/Photobleaching">Photobleaching</a>
 
       <a href="/Team:IIT_Delhi/Photobleaching">Photobleaching</a>
 
       <a href="/Team:IIT_Delhi/Promoter">Promoter strength</a>
 
       <a href="/Team:IIT_Delhi/Promoter">Promoter strength</a>
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20/4<br><ul>
 
<li>First team meet up
 
<li>Introduction of team members</ul><br>
 
25/4<br><ul>
 
<li> Discussion on DNA structure and replication, Chargaff’s rule, primers and enzymes</ul><br>
 
27/4<br><ul>
 
<li> Discussion on plasmids, digestion, ligation, transformation and 3A assembly
 
<li> Brainstorming on this year’s project - square wave generator</ul><br>
 
28/4<br><ul>
 
<li>Discussion on reference papers of repressilator, synchronous oscillations,
 
robustness etc.</ul><br>
 
29/4<br><ul>
 
<li> Discussion on Gel electrophoresis and blue-white screening</ul><br>
 
  
</h2>
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       <h2  class="boogaloo_font "><span style="font-size:30px;line-height:30px;cursor:pointer;display:block">We planned early on to document the work we did and efforts we put into our project, on a daily basis,hence, we were able to compile a comprehensive notebook accounting for all the events that took place.<br><br>April</span></h2>
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      <h2  class="boogaloo_font "><span style="font-size:30px;cursor:pointer">May</span></h2>
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01/05<br>
 
First lab visit of the team and instructions on how to handle the instruments and
 
safety guidelines were presented<br>
 
02/05<br>
 
<ol>
 
<li> Practicing inoculation, streaking and spreading of bacterial cultures of a biobrick used
 
in previous year
 
<li> LA plates and LB media preparation</ol><br>
 
03/05<br>
 
<ol>
 
<li> Practicing plasmid isolation and transformation with the part inoculated yesterday
 
<li> Inoculation of following parts from iGEM kit plate:-
 
<ul>
 
<li> RBS + GFP + T(without deg tag)
 
<li> RBS + TetR + T
 
<li>SYFP2
 
<li> J23119 promoter
 
<li> J23107 promoter</ul><br>
 
<li> Plasmid isolation of :-<br>
 
<ul>
 
<li> RBS + GFP + T(without deg tag)
 
<li> RBS + TetR + T
 
<li> SYFP2
 
<li> J23119 promoter
 
<li> J23107 promoter</ul><br>
 
<li> Transformation of plasmids mentioned previously following iGEM protocol</ol><br>
 
04/05<br>
 
<ol>
 
<li> Discussion on toggles switches and oscillators in relation to iGEM IITD project - 2016.
 
More detailed discussion on light activated self-repression and its effect on oscillator
 
frequency.
 
<li> Discussion on IITD 2015 project
 
<li> Discussion on some iGEM projects of other countries.
 
<li> Received the colonies transformed yesterday</ol><br>
 
06/05<br>
 
Further discussion on the research papers and different aspects of project<br>
 
09/05<br>
 
Formation of a marketing and human practices team and discussion on marketing
 
strategies used last year.<br>
 
11/05<br>
 
<ol>
 
<li>Video and content for crowd-funding initiative
 
<li>Mails and calls to potential sponsors</ol><br>
 
13/05<br>
 
<ol><li>Discussion on gene regulation in prokaryotes and eukaryotes with special emphasis on
 
prokaryotes.
 
<li> Discussion on how we can modify the Lac system by keeping its switching on properties
 
intact and replacing the gene part with, suppose, insulin such that presence Lactose
 
molecules trigger the activation of insulin gene.
 
<li> Discussion on the different class of promoters.</ol><br>
 
15/05<br>
 
<ol>
 
<li> Discussion on PCR
 
<li> Sample PCR product formed and ran on gel.</ol><br>
 
17/05
 
 1. Logic Gates:
 
a) AND GATE:
 
 
 discussion on simple AND gate
 
 AND gate with modified tRNA (Discussion on tRNA that would read TAG as Serine)
 
b) OR GATE
 
c) NOR GATE
 
d) XOR GATE: Using combination of different gates i.e. OR , AND , NOR, NAND, NOT and
 
further discussion on other ways of XOR gate formation.
 
 2. Steady state: Rate of accumulation=0
 
 3. Discussion on different equations for rate of change of protein and rate of change of RNA
 
(First and second order reactions) by discussing the journey from promoter to mRNA to
 
protein to nothingness.
 
 4. Enzyme Kinetics
 
 S+E === ES ----> P+E
 
 Differential equations of rate of change of ES and P discussed with equations of conservation
 
of mass for the same.
 
 Graphical analysis of ES/E0 vs S/S0 with discussion on quasi steady state.
 
 Graphical discussion on Concentration vs Time of the reaction.
 
 5. Michaelis menten equation
 
 6. Discussion on Degradation Tag and how it works in a bacterium where translation and
 
transcription happen in the same compartment and there are no stop codons.
 
 7. Discussion on Cancer Cell about how it was thought to find a cure of cancer using
 
these logic gates with inputs as miRNA(regulate genes) and output as apoptosis.
 
 8. Example of virus that can be used as vectors for human cells. e.g. Vaccinia virus
 
18/05
 
 Discussion on ways to generate different signal responses in bacterial gene regulatory
 
circuits
 
22/05
 
 Discussion on :-
 
1) Chemotactic response in bacteria
 
2) Temporal and spatial response in chemotaxis
 
3) Mechanism of chemotaxis in bacteria flagellum
 
4) Electrical model of chemotaxis
 
5) Adaptation time of chemotaxis
 
6) Finding electrical analogue of the biological system by varying frequency of the electrical circuit
 
made
 
7) Finding optimal frequency
 
8) Biological Robustness and causes of it in a biological system
 
9) System controls, bistability, modularity, buffering, bow-tie framework, decoupling
 
10) Robustness in signal transduction pathways and insect's segmental development
 
11) Robustness trade-offs
 
24/05
 
 Discussion on :-
 
1) Design principles of biochemical oscillators
 
2) Time delay's relevance in oscillation
 
3) Introduction of time delay with Intermediates
 
4) Limit cycles, conservative cycles and circadian rhythms
 
5) Time delay by positive feedback
 
6) Different classes of feedback
 
7) Linear and hyperbolic response
 
8) Goldbeter Kosher Function
 
9) Sniffers
 
10) Toggle Switch
 
 
11) Mutual inhibition and mutual activation
 
26/05
 
 Discussion on :-
 
1) A brief review of Sniffers, Buzzers and Toggle switch
 
2) Negative feedback oscillations
 
3) Positive feedback oscillations
 
4) Neutral feedback oscillations
 
5) Sub-critical and Super-critical hop bit
 
6) Study of mitosis
 
27/05
 
 Prepared lab supplies like LA, LB etc. to be used in the lab
 
29/05
 
1) Preparation of LuxI and PLux double digest using restriction enzymes EcoRI and PstI
 
2) Preparation of buffer and electrophoresis gel
 
3) Loading ladder, control and digest in the wells in gel
 
4) Electrophoresis and subsequent observation of gel to ensure digestion of plasmids
 
31/05
 
Discussion on:-
 
1) Plasmid isolation
 
2) Lysis solutions and their composition
 
3) Function of each component in the solutions
 
 
</h2>
 
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      <h2  class="boogaloo_font "><span style="font-size:30px;cursor:pointer" onclick="openNav()">May</span></h2>
 
 
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Latest revision as of 22:19, 1 November 2017

iGEM IIT Delhi

We planned early on to document the work we did and efforts we put into our project, on a daily basis,hence, we were able to compile a comprehensive notebook accounting for all the events that took place.

April

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May

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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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