Difference between revisions of "GDSYZX-United/PROJECT/preparation arabidopsis protoplast"

 
(6 intermediate revisions by 2 users not shown)
Line 24: Line 24:
 
nav {padding-top:10px;}
 
nav {padding-top:10px;}
 
.text_font {
 
.text_font {
 +
                        text-indent:1em;
 
line-height:120%;
 
line-height:120%;
 
font-size:130%
 
font-size:130%
Line 39: Line 40:
 
<ul class="nav navbar-nav navbar-right">
 
<ul class="nav navbar-nav navbar-right">
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United"> HOME </a>
+
<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United">HOME</a>
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> TEAM <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">TEAM<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
 
<li><a href="https://2016.igem.org/GDSYZX-United/TEAM/team_members">Team members</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/TEAM/team_members">Team members</a></li>
 +
<li><a href="https://2016.igem.org/Team:GDSYZX-United/Collaborations">Collarboration</a></li>
 +
<li><a href="https://2016.igem.org/Team:GDSYZX-United/Attributions">Attribution</a></li>
 
</ul>
 
</ul>
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> PROJECT <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">PROJECT<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
−
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/project_description">Project description</a></li>
+
<li><a href="https://2016.igem.org/Team:GDSYZX-United/Description">Project description</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/preparation_arabidopsis_protoplast">Preparation of the Arabidopsis protoplast</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/preparation_arabidopsis_protoplast">Preparation of the Arabidopsis protoplast</a></li>
−
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/result">Result</a></li>
+
<li><a href="https://2016.igem.org/Team:GDSYZX-United/Proof">Proof</a></li>
 +
<li><a href="https://2016.igem.org/Team:GDSYZX-United/Demonstrate">Demonstrate</a></li>
 
</ul>
 
</ul>
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> PARTS <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United/results">RESULTS</a>
 +
</li>
 +
<li class="dropdown">
 +
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">PARTS<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
 
<li><a href="https://2016.igem.org/GDSYZX-United/PARTS/parts">Parts</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PARTS/parts">Parts</a></li>
Line 64: Line 71:
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> NOTEBOOK <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">NOTEBOOK<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
 
    <li><a href="https://2016.igem.org/GDSYZX-United/NOTEBOOK/day_note">Day notes</a></li>
 
    <li><a href="https://2016.igem.org/GDSYZX-United/NOTEBOOK/day_note">Day notes</a></li>
Line 72: Line 79:
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> HUMAN PRACTICE <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">HUMAN PRACTICE<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
−
<li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/human_practice">Human Practice</a></li>
+
<li><a href="https://2016.igem.org/Team:GDSYZX-United/HP/Silver">Silver</a></li>
 +
<li><a href="https://2016.igem.org/Team:GDSYZX-United/HP/Gold">Gold</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/album">Album</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/album">Album</a></li>
 
</ul>
 
</ul>
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> SAFETY <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown">SAFETY<span class="caret"></span></a>
 
<ul role="menu" class="dropdown-menu">
 
<ul role="menu" class="dropdown-menu">
 
<li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/lab_safety">Lab safety</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/lab_safety">Lab safety</a></li>
Line 86: Line 94:
 
</li>
 
</li>
 
<li class="dropdown">
 
<li class="dropdown">
−
<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> ENTREPRENEURSHIP <span class="caret"></span></a>
+
<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United/Entrepreneurship">ENTREPRENEURSHIP</a>
−
<ul role="menu" class="dropdown-menu">
+
−
<li><a href="#">1</a></li>
+
−
<li><a href="#">2</a></li>
+
−
<li><a href="#">3</a></li>
+
−
<li><a href="#">4</a></li>
+
−
</ul>
+
 
</li>
 
</li>
 
</ul>
 
</ul>
Line 113: Line 115:
 
<div class="col-sm-2"></div>
 
<div class="col-sm-2"></div>
 
<div class="col-sm-8 content">
 
<div class="col-sm-8 content">
−
<p><div class="text_font"><strong>Place:</strong>Biology Experiment Teaching Center of Sun Yat-sen University</div></p>
+
<p><div class="text_font" style="text-indent:0em;"><strong>Place:</strong>Biology Experiment Teaching Center of Sun Yat-sen University</div></p>
−
<p><div class="text_font"><strong>Date:</strong>July 28th, 2016</div></p>
+
 
<p><div class="text_font"><strong>Experimental apparatus:</strong>Shaking table, 50 ml centrifuge tube, filtrating equipment, analytical balance, measuring cylinder</div></p>
 
<p><div class="text_font"><strong>Experimental apparatus:</strong>Shaking table, 50 ml centrifuge tube, filtrating equipment, analytical balance, measuring cylinder</div></p>
 
<p><div class="text_font"><strong>Experimental drugs:</strong>Cellulase hydrolysate, W5 solution</div></p>
 
<p><div class="text_font"><strong>Experimental drugs:</strong>Cellulase hydrolysate, W5 solution</div></p>
Line 126: Line 127:
 
<p><div class="text_font"><strong>Procedure:</strong></div></p>
 
<p><div class="text_font"><strong>Procedure:</strong></div></p>
 
<ol>
 
<ol>
−
<p><div class="text_font"><li>
+
<p><div class="text_font"><li style="text-indent:0em;">
 
the configuration of Cellulase hydrolysate<br>
 
the configuration of Cellulase hydrolysate<br>
 
<table class="table">
 
<table class="table">
Line 175: Line 176:
 
</table>
 
</table>
 
</li></div></p>
 
</li></div></p>
−
<p><div class="text_font"><li>
+
<p><div class="text_font"><li style="text-indent:0em;">
 
Pick Arabidopsis leaves and cut out them with 0.5-1mm, then immerse it in 5ml  Cellulase hydrolysate.
 
Pick Arabidopsis leaves and cut out them with 0.5-1mm, then immerse it in 5ml  Cellulase hydrolysate.
 
</li></div></p>
 
</li></div></p>
−
<p><div class="text_font"><li>
+
<p><div class="text_font"><li style="text-indent:0em;">
 
Put them in shaking table for 30min in the dark, then waiting for another 3h without shaking. When it comes to green and then gently shake it.
 
Put them in shaking table for 30min in the dark, then waiting for another 3h without shaking. When it comes to green and then gently shake it.
 
</li></div></p>
 
</li></div></p>
−
<p><div class="text_font"><li>
+
<p><div class="text_font"><li style="text-indent:0em;">
 
(Adding 5ml W5 solution and then filtrating.)
 
(Adding 5ml W5 solution and then filtrating.)
 
</li></div></p>
 
</li></div></p>
−
<p><div class="text_font"><li>
+
<p><div class="text_font"><li style="text-indent:0em;">
 
The protoplast was observed under the microscope
 
The protoplast was observed under the microscope
 
</li></div></p>
 
</li></div></p>
Line 195: Line 196:
 
<div class="col-sm-2"></div>
 
<div class="col-sm-2"></div>
 
<div class="col-sm-8 content">
 
<div class="col-sm-8 content">
−
<p><div class="text_font"><strong>Result:</strong></div></p>
+
−
<p><div class="text_font">There is no protoplast in Cellulase hydrolysate (it didn’t come to green), and the cell wall of young leaves were not damaged.</div></p>
+
 
<p><div class="text_font"><strong>Introspection:</strong></div></p>
 
<p><div class="text_font"><strong>Introspection:</strong></div></p>
 
<p><div class="text_font">
 
<p><div class="text_font">
 
<ol>
 
<ol>
−
<li>Cutting leaves without using the blade but by hand , or deal it with a scalpel pulling, which may damaged the cells.</li>
+
<li style="text-indent:0em;">Cutting leaves without using the blade but by hand , or deal it with a scalpel pulling, which may damaged the cells.</li>
−
<li>The shaking table should not exceed 80r/min.</li>
+
<li style="text-indent:0em;">The shaking table should not exceed 80r/min.</li>
 
</ol>
 
</ol>
 
</div></p>
 
</div></p>

Latest revision as of 14:53, 17 October 2016

Preparation of the Arabidopsis protoplast

Place:Biology Experiment Teaching Center of Sun Yat-sen University

Experimental apparatus:Shaking table, 50 ml centrifuge tube, filtrating equipment, analytical balance, measuring cylinder

Experimental drugs:Cellulase hydrolysate, W5 solution

Procedure:

  1. the configuration of Cellulase hydrolysate
    Cellulase 0.225g
    Mecerozyme R10 0.045g
    mannitol 1.09g
    KCl 1.5g
    MES 4.18g
    water 12ml
    cooled down to room temperature after waiting in 55℃ water 10min.
    CaCl2 1.11g
    BSA 1ml
    water 2ml
    filtrating

  2. Pick Arabidopsis leaves and cut out them with 0.5-1mm, then immerse it in 5ml Cellulase hydrolysate.

  3. Put them in shaking table for 30min in the dark, then waiting for another 3h without shaking. When it comes to green and then gently shake it.

  4. (Adding 5ml W5 solution and then filtrating.)

  5. The protoplast was observed under the microscope

Introspection:

  1. Cutting leaves without using the blade but by hand , or deal it with a scalpel pulling, which may damaged the cells.
  2. The shaking table should not exceed 80r/min.

image

Filtrate during the configuration of Cellulase hydrolysate

image

Cut out the leaves

image

Immerse Arabidopsis leaves in 5ml Cellulase hydrolysate.

image

Prepare the microscopic slides

image

Cellulase hydrolysate under microscope (nothing).

image

Plant cells under microscope.