Difference between revisions of "GDSYZX-United/PROJECT/preparation arabidopsis protoplast"
Thomasboboto (Talk | contribs) (Created page with "<html> <head> <meta charset="utf-8"> <meta name="viewport" content="width=device-width, initial-scale=1.0"> <link href="https://2016.igem.org/Template:GDSYZX-Uni...") |
Thomasboboto (Talk | contribs) |
||
| Line 213: | Line 213: | ||
<div class="col-sm-8"> | <div class="col-sm-8"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/a/a6/T--GDSYZX-United--project_prepration1.jpg"> |
<p><div class="text_font">Filtrate during the configuration of Cellulase hydrolysate</div></p> | <p><div class="text_font">Filtrate during the configuration of Cellulase hydrolysate</div></p> | ||
</div> | </div> | ||
| Line 219: | Line 219: | ||
<div class="col-sm-4"> | <div class="col-sm-4"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/d/da/T--GDSYZX-United--project_prepration2.jpg"> |
<p><div class="text_font">Cut out the leaves</div></p> | <p><div class="text_font">Cut out the leaves</div></p> | ||
</div> | </div> | ||
| Line 232: | Line 232: | ||
<div class="col-sm-4"> | <div class="col-sm-4"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/c/cf/T--GDSYZX-United--project_prepration3.jpg"> |
<p><div class="text_font">Immerse Arabidopsis leaves in 5ml Cellulase hydrolysate.</div></p> | <p><div class="text_font">Immerse Arabidopsis leaves in 5ml Cellulase hydrolysate.</div></p> | ||
</div> | </div> | ||
| Line 238: | Line 238: | ||
<div class="col-sm-8"> | <div class="col-sm-8"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/e/e8/T--GDSYZX-United--project_prepration4.jpg"> |
<p><div class="text_font">Prepare the microscopic slides </div></p> | <p><div class="text_font">Prepare the microscopic slides </div></p> | ||
</div> | </div> | ||
| Line 251: | Line 251: | ||
<div class="col-sm-6"> | <div class="col-sm-6"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/2/2f/T--GDSYZX-United--project_prepration5.jpg"> |
<p><div class="text_font">Cellulase hydrolysate under microscope (nothing).</div></p> | <p><div class="text_font">Cellulase hydrolysate under microscope (nothing).</div></p> | ||
</div> | </div> | ||
| Line 257: | Line 257: | ||
<div class="col-sm-6"> | <div class="col-sm-6"> | ||
<div align="center"> | <div align="center"> | ||
| − | <img alt="image" class="img-responsive" src=" | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/c/c8/T--GDSYZX-United--project_prepration6.jpg"> |
<p><div class="text_font">Plant cells under microscope.</div></p> | <p><div class="text_font">Plant cells under microscope.</div></p> | ||
</div> | </div> | ||
Revision as of 17:40, 1 October 2016
Preparation of the Arabidopsis protoplast
Place:Biology Experiment Teaching Center of Sun Yat-sen University
Date:July 28th, 2016
Experimental apparatus:Shaking table, 50 ml centrifuge tube, filtrating equipment, analytical balance, measuring cylinder
Experimental drugs:Cellulase hydrolysate, W5 solution
Procedure:
-
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 - Pick Arabidopsis leaves and cut out them with 0.5-1mm, then immerse it in 5ml Cellulase hydrolysate.
- 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.
- (Adding 5ml W5 solution and then filtrating.)
- The protoplast was observed under the microscope
Result:
There is no protoplast in Cellulase hydrolysate (it didn’t come to green), and the cell wall of young leaves were not damaged.
Introspection:
- Cutting leaves without using the blade but by hand , or deal it with a scalpel pulling, which may damaged the cells.
- The shaking table should not exceed 80r/min.
Filtrate during the configuration of Cellulase hydrolysate
Cut out the leaves
Immerse Arabidopsis leaves in 5ml Cellulase hydrolysate.
Prepare the microscopic slides
Cellulase hydrolysate under microscope (nothing).
Plant cells under microscope.
