Team:Groningen/protocols

From 2013.igem.org

(Difference between revisions)
 
(31 intermediate revisions not shown)
Line 1: Line 1:
-
'''B. subtilis transformation'''
+
<html><!--
-
<br/>Losick protocol
+
Just put the link between the brackets as below. The link should look like: :Team:Groningen/protocols/"protocol-name" Try to think of a name that makes sense, so please don't put any bullshit there! Anyway after the "|" you can type the name of your link. Please use the full protocol name, so it is clear for anyway it is about.
 +
I've added the headers as an example, please change to something that makes more sense and feel free to add more
 +
-->
 +
<h1>protocols</h1>
-
<br/>-1 Day: Streak out strain and incubate plate o/n at 37 °C.
+
</html>
 +
<h2>General protocols</h2>
 +
<ul>
 +
<li>[[Team:Groningen/protocols/GelElectrophoresis | Gel electrophoresis]]</li>
 +
<li>[[Team:Groningen/protocols/PCR | PCR]]</li>
 +
<li>[[Team:Groningen/protocols/Colony PCR |Colony PCR]]</li>
 +
<li>[[Team:Groningen/protocols/Digestion | Restriction digestion protocol]]</li>
 +
<li>[[Team:Groningen/protocols/Ligation | Ligation protocol]]</li>
 +
</ul>
 +
<br>
 +
<h2>''Escherichia coli'' protocols</h2>
-
<br/>Transformation (D-Day):
+
*[[:Team:Groningen/protocols/Transformation_EC | ''E. coli'' transformation protocol]]
-
 
+
<br>
-
<br/>1. Pick up a nice big colony and drop it in 2 ml of completed MC (1x)(see sub1).
+
<h2>''Bacillus subtilis'' protocols</h2>
-
<br/>2. Grow at 37 °C for 5 hours (more if culture is not really turbid).
+
*[[:Team:Groningen/protocols/Genomic DNA extraction | Genomic DNA extraction of ''B. subtilis'']]
-
<br/>3. Mix 400 µl of culture with DNA* in fresh tube (i.e. 15 ml tubes loosely closed – aeration) (*usually 1 µl. Then 10 µl of Quiagen plasmid miniprep or <1 µl of  chromosal prep)
+
*[[:Team:Groningen/protocols/Transformation | ''B. subtilis'' transformation protocol]]
-
<br/>4. Grow for an additional 2 hours at 37 °C.
+
*[[:Team:Groningen/protocols/biofilm_media | Biofilm formation protocol]]
-
<br/>5. Plate all on selective antibiotic plates, and incubates at 37°C o/n.
+
*[[:Team:Groningen/protocols/Motility_assay | Motility assay]]
-
 
+
-
 
+
-
<br/>Sub1: Copmpetence medium (MC completed)
+
-
<table border="1">
+
-
<tr>
+
-
<td>H20</td>
+
-
<td>1,8 ml</td>
+
-
<td></td>
+
-
</tr>
+
-
<tr>
+
-
<td>10x MC (sub2)</td>
+
-
<td>200 ul</td>
+
-
<td> filter sterilize</td>
+
-
</tr>
+
-
<tr>
+
-
<td>MgSO4</td>
+
-
<td>6,7 ul</td>
+
-
<td>autoclave sterilize</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Tryptophan 1%</td>
+
-
<td>10 ul</td>
+
-
<td>filter steralize, wrap in Al</td>
+
-
</tr></table>
+
-
 
+
-
<br/>Sub2: MC 10x
+
-
<table border="1">
+
-
<tr>
+
-
<td>for</td>
+
-
<td>100 ml</td>
+
-
<td>10 ml</td>
+
-
</tr>
+
-
<tr>
+
-
<td>K2H PO4</td>
+
-
<td>14,036g</td>
+
-
<td> 1,4036g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>KH2 PO$</td>
+
-
<td>5,239g</td>
+
-
<td>0,5239g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Glucose</td>
+
-
<td>20g</td>
+
-
<td>2g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Tri-Na Citrate 300mM(Sub3)</td>
+
-
<td>10ml</td>
+
-
<td>1ml</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Ferric NH4 citrate(Sub4)</td>
+
-
<td>1ml</td>
+
-
<td>0,1ml</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Casein Hydrolysate</td>
+
-
<td>1g</td>
+
-
<td>0,1g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Potassium Glutamate</td>
+
-
<td>2g</td>
+
-
<td>0,2g</td>
+
-
</tr>
+
-
</table>
+
-
Add 50ml H2O, Mix, add H2O till 100ml, Filter sterilize and freeze at -20 °C
+
-
 
+
-
 
+
-
<br/>Sub3: Tri-Na Citrate 300mM
+
-
<table border="1">
+
-
<tr>
+
-
<td>Tri-Na Citrate</td>
+
-
<td>0,8823g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>H2O</td>
+
-
<td>10ml</td>
+
-
</tr>
+
-
</table>
+
-
Wrap in aluminum
+
-
 
+
-
<br/>Sub4: Ferric NH4 citrate
+
-
<table border="1">
+
-
<tr>
+
-
<td>Ferric NH4 citrate</td>
+
-
<td>0,22g</td>
+
-
</tr>
+
-
<tr>
+
-
<td>H2O</td>
+
-
<td>10ml</td>
+
-
</tr>
+
-
</table>
+
-
Wrap in aluminum
+

Latest revision as of 02:26, 5 October 2013

protocols

General protocols


Escherichia coli protocols


Bacillus subtilis protocols

Retrieved from "http://2013.igem.org/Team:Groningen/protocols"