Difference between revisions of "Team:IISER Kolkata/Experiments"

 
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<h1 class="heading">Experiments</h1>
  
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<section class="sec white">
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<p>A great man once said, <i>&quot;The true method of knowledge is through experiment.&quot;</i> And in the quest to gain knowledge we did a lot of experiments.</p>
 +
<p>Most of our work dealt with genetically engineering our bacterial chassis i.e. <i>E. coli</i>. Additionally, to support our model and to gain some meaningful insights into the system, we did a variety of experiment involving Macrophage and <i>Leishmania major</i>.</p>
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</section>
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<section class="sec white">
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<h3>Bacterial Protocols</h3>
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<div class="exp-list">
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<a href="https://static.igem.org/mediawiki/2019/b/b9/T--IISER_Kolkata--1_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>1</h1></div>
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<h5>LB Media Preparation</h5>
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</div></a>
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<a href="https://static.igem.org/mediawiki/2019/5/50/T--IISER_Kolkata--2_bac.pdf" target="_blank"><div class="protocol">
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<div class="index"><h1>2</h1></div>
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<h5>SOB Media Preparation</h5>
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</div></a>
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<a href="https://static.igem.org/mediawiki/2019/8/8d/T--IISER_Kolkata--3_bac.pdf" target="_blank"><div class="protocol">
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<div class="index"><h1>3</h1></div>
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<h5>50X TAE Buffer</h5>
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</div></a>
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<a href="https://static.igem.org/mediawiki/2019/b/bd/T--IISER_Kolkata--4_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>4</h1></div>
 +
<h5>CCMB Buffer Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/f/fd/T--IISER_Kolkata--5_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>5</h1></div>
 +
<h5>Calcium chloride Competent Cell Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/4/45/T--IISER_Kolkata--6_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>6</h1></div>
 +
<h5>Hanahan Competent Cell Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/5/52/T--IISER_Kolkata--7_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>7</h1></div>
 +
<h5>Electrocompetent Cell Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/f/f5/T--IISER_Kolkata--8_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>8</h1></div>
 +
<h5>Heat Shock Transformation of Chemically Competent Cells</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/a/ac/T--IISER_Kolkata--9_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>9</h1></div>
 +
<h5>Transformation by Electroporation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/2/23/T--IISER_Kolkata--10_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>10</h1></div>
 +
<h5>PCR Amplification: Q5® H-F MM</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/2/28/T--IISER_Kolkata--11_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>11</h1></div>
 +
<h5>PCR Amplification: Taq Pol</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/1/16/T--IISER_Kolkata--12_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>12</h1></div>
 +
<h5>Restriction Digestion</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/4/40/T--IISER_Kolkata--13_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>13</h1></div>
 +
<h5>Ligation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/a/aa/T--IISER_Kolkata--14_bac.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>14</h1></div>
 +
<h5>SDS-PAGE Electrophoresis</h5>
 +
</div></a>
 +
</div>
 +
</section>
 +
 +
<section class="sec white">
 +
<h3><i>Leishmania major</i> culture protocols</h3>
 +
<div class="exp-list">
 +
<a href="https://static.igem.org/mediawiki/2019/2/25/T--IISER_Kolkata--cell_1.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>1</h1></div>
 +
<h5>M199 Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/b/b0/T--IISER_Kolkata--cell_2.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>2</h1></div>
 +
<h5>M199 Media with different Hemin Concentrations</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/f/ff/T--IISER_Kolkata--cell_3.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>3</h1></div>
 +
<h5>Culturing <i>Leishmania major</i></h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/2/23/T--IISER_Kolkata--cell_4.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>4</h1></div>
 +
<h5><i>Leishmania</i>-Macrophage Infection</h5>
 +
</div></a>
 +
</div>
 +
</section>
 +
 +
<section class="sec white">
 +
<h3>Macrophage culture protocols</h3>
 +
<div class="exp-list">
 +
<a href="https://static.igem.org/mediawiki/2019/a/a2/T--IISER_Kolkata--cell_5.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>1</h1></div>
 +
<h5>DMEM Culture Media Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/9/90/T--IISER_Kolkata--cell_6.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>2</h1></div>
 +
<h5>Phosphate Buffer Saline Preparation</h5>
 +
</div></a>
 +
<a href="https://static.igem.org/mediawiki/2019/d/d9/T--IISER_Kolkata--cell_7.pdf" target="_blank"><div class="protocol">
 +
<div class="index"><h1>3</h1></div>
 +
<h5>Murine Macrophage J774A.1 Culturing</h5>
 +
</div></a>
 +
</div>
 +
</section>
  
 +
</div>
 
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Latest revision as of 02:17, 22 October 2019

Experiments

A great man once said, "The true method of knowledge is through experiment." And in the quest to gain knowledge we did a lot of experiments.

Most of our work dealt with genetically engineering our bacterial chassis i.e. E. coli. Additionally, to support our model and to gain some meaningful insights into the system, we did a variety of experiment involving Macrophage and Leishmania major.

Bacterial Protocols

1

LB Media Preparation

2

SOB Media Preparation

3

50X TAE Buffer

4

CCMB Buffer Preparation

5

Calcium chloride Competent Cell Preparation

6

Hanahan Competent Cell Preparation

7

Electrocompetent Cell Preparation

8

Heat Shock Transformation of Chemically Competent Cells

9

Transformation by Electroporation

10

PCR Amplification: Q5® H-F MM

11

PCR Amplification: Taq Pol

12

Restriction Digestion

13

Ligation

14

SDS-PAGE Electrophoresis

Leishmania major culture protocols

1

M199 Preparation

2

M199 Media with different Hemin Concentrations

3

Culturing Leishmania major

4

Leishmania-Macrophage Infection

Macrophage culture protocols

1

DMEM Culture Media Preparation

2

Phosphate Buffer Saline Preparation

3

Murine Macrophage J774A.1 Culturing