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Team:Imperial College London/Protocols Chemotaxis
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<h1>Phyto-Route</h1> | <h1>Phyto-Route</h1> | ||
| - | <h2>Tryptone | + | <h2>Tryptone broth</h2> |
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
<h2>PBS</h2> | <h2>PBS</h2> | ||
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| - | <h2>Motility | + | <h2>Motility medium</h2> |
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
<h2>Preparation</h2> | <h2>Preparation</h2> | ||
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| - | <h2>Agar | + | <h2>Agar plugin</h2> |
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| - | <h2>Semi-solid | + | <h2>Semi-solid agar</h2> |
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| - | <h2>Capillary | + | <h2>Capillary assay</h2> |
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
<h2>Plant uptake</h2> | <h2>Plant uptake</h2> | ||
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| - | <p>- | + | <p>- Grow GFP-expressing <i>E. coli</i> to exponential phase<br> |
| - | - | + | - Spin down bacteria (5000 rpm for 10 min) and remove LB medium supernatant.<br> |
| - | - | + | - Wash twice with wash buffer (5 mM MES).<br> |
| - | - | + | - Re-suspend in wash buffer so that the bacteria are at OD 30. <br> |
| - | - | + | - Put 10 Arabidopsis seedlings into 100 ml of growth media each. <br> |
| - | - | + | - Add bacteria to plant growth media, add the same amount of wash buffer to the negative control. <br> |
| - | - | + | - Image after 12 h and 24 h.</p> |
<p><b>Some notes</b><br> | <p><b>Some notes</b><br> | ||
| - | - Some departments have very strong restrictions on bringing bacteria into dedicated plant growing areas<br> | + | - Some departments have very strong restrictions on bringing bacteria into dedicated plant growing areas. <br> |
- After talking to our health and safety officers, we decided to do all preparation and disposal of bacterial cultures as well as addition of the bacteria to plant media in biosafety hoods in dedicated level 1 or 2 laboratories so that there was no danger of dispersing the bacteria around the plant rooms.</p> | - After talking to our health and safety officers, we decided to do all preparation and disposal of bacterial cultures as well as addition of the bacteria to plant media in biosafety hoods in dedicated level 1 or 2 laboratories so that there was no danger of dispersing the bacteria around the plant rooms.</p> | ||
</body> | </body> | ||
</html> | </html> | ||
Revision as of 16:18, 18 September 2011
Protocols
This page lists all the protocols used in our project. We have classified them into five main categories as follow.
Phyto-Route
Tryptone broth
PBS
Motility medium
Preparation
Agar plugin
Semi-solid agar
Capillary assay
Plant uptake
- Grow GFP-expressing E. coli to exponential phase
- Spin down bacteria (5000 rpm for 10 min) and remove LB medium supernatant.
- Wash twice with wash buffer (5 mM MES).
- Re-suspend in wash buffer so that the bacteria are at OD 30.
- Put 10 Arabidopsis seedlings into 100 ml of growth media each.
- Add bacteria to plant growth media, add the same amount of wash buffer to the negative control.
- Image after 12 h and 24 h.
Some notes
- Some departments have very strong restrictions on bringing bacteria into dedicated plant growing areas.
- After talking to our health and safety officers, we decided to do all preparation and disposal of bacterial cultures as well as addition of the bacteria to plant media in biosafety hoods in dedicated level 1 or 2 laboratories so that there was no danger of dispersing the bacteria around the plant rooms.
