Team:Cambridge/Bacillus subtilis transformation
From 2008.igem.org
(One intermediate revision not shown) | |||
Line 11: | Line 11: | ||
class="b4f"></b> | class="b4f"></b> | ||
<div class="contentf"> | <div class="contentf"> | ||
- | <div style="height: 400; background: white; line-height:170% padding: | + | <div style="height: 400; background: white; line-height:170% padding:5px;"> |
- | 5px;"> | + | |
<div style="color: white; font: 2px;">x</div> | <div style="color: white; font: 2px;">x</div> | ||
</html> | </html> | ||
Line 20: | Line 19: | ||
==Our aim== | ==Our aim== | ||
- | We want to transform ''Bacillus subtilis'' with two kinds of vectors: episomal vectors and integration vector. We would like to obtain good efficiency, no contamination and some precise tests confirm our transformations and to be able to standardize our protocol | + | We want to transform ''Bacillus subtilis'' with two kinds of vectors: episomal vectors and integration vector. We would like to obtain good efficiency, no contamination and some precise tests confirm our transformations and to be able to standardize our protocol. This protocol is modified from the one provided by the Bacillus Genetic Stock Center (http://www.bgsc.org/catalog.htm): we have made the addition of tryptophan, and experimented with different sterile methods for preparation of the growth media. |
==Material== | ==Material== | ||
Line 143: | Line 142: | ||
* Transformation in IA751 : Amylase test | * Transformation in IA751 : Amylase test | ||
[[Image: Protocol13.jpg | thumb | 300px | center | IA751 transformed with ECE112 : SUCCESSFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]] | [[Image: Protocol13.jpg | thumb | 300px | center | IA751 transformed with ECE112 : SUCCESSFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]] | ||
+ | |||
+ | <html> | ||
+ | <div style="color: white; font: 2px;">x</div> | ||
+ | </div></div> | ||
+ | <b class="b4f"></b><b class="b3f"></b><b class="b2f"></b><b class="b1f"></b> | ||
+ | </td></tr></table></table> | ||
+ | </html> | ||
+ | |||
+ | __NOEDITSECTION__ |
Latest revision as of 20:39, 29 October 2008
x
Transformation of Bacillus subtilisOur aimWe want to transform Bacillus subtilis with two kinds of vectors: episomal vectors and integration vector. We would like to obtain good efficiency, no contamination and some precise tests confirm our transformations and to be able to standardize our protocol. This protocol is modified from the one provided by the Bacillus Genetic Stock Center (http://www.bgsc.org/catalog.htm): we have made the addition of tryptophan, and experimented with different sterile methods for preparation of the growth media. Material
Media Preparation10X Medium A base:
10X Bacillus salts:
Medium A
Medium B
Important:
ProtocolsMaking Bacillus competent
Transforming
Glycerol stocks
ResultsTransformation with episomal vectorsECE166
We obtained consistent good results with this transformation despite the low efficiency.
Transformation with integration vectorsECE153
Protocol of the Amylase Test:
Protocol of the Xylose Test:
We obtain good result for transformation. Amylase test is positive, but we have not manage to induce Pxyl. ECE112
x
|