Team:Groningen/protocols

From 2013.igem.org

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'''B. subtilis transformation'''
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<html><!--
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<br/>Losick protocol
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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.
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I've added the headers as an example, please change to something that makes more sense and feel free to add more
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<h1>protocols</h1>
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<br/>-1 Day: Streak out strain and incubate plate o/n at 37 °C.
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</html>
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<h2>General protocols</h2>
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<ul>
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<li>[[Team:Groningen/protocols/GelElectrophoresis | Gel electrophoresis]]</li>
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<li>[[Team:Groningen/protocols/PCR | PCR]]</li>
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<li>[[Team:Groningen/protocols/Colony PCR |Colony PCR]]</li>
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<li>[[Team:Groningen/protocols/Digestion | Restriction digestion protocol]]</li>
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<li>[[Team:Groningen/protocols/Ligation | Ligation protocol]]</li>
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</ul>
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<br>
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<h2>''Escherichia coli'' protocols</h2>
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<br/>Transformation (D-Day):
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*[[:Team:Groningen/protocols/Transformation_EC | ''E. coli'' transformation protocol]]
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<br>
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<br/>1. Pick up a nice big colony and drop it in 2 ml of completed MC (1x)(see sub1).
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<h2>''Bacillus subtilis'' protocols</h2>
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<br/>2. Grow at 37 °C for 5 hours (more if culture is not really turbid).
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*[[:Team:Groningen/protocols/Genomic DNA extraction | Genomic DNA extraction of ''B. subtilis'']]
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<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)
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*[[:Team:Groningen/protocols/Transformation | ''B. subtilis'' transformation protocol]]
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<br/>4. Grow for an additional 2 hours at 37 °C.
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*[[:Team:Groningen/protocols/biofilm_media | Biofilm formation protocol]]
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<br/>5. Plate all on selective antibiotic plates, and incubates at 37°C o/n.
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*[[:Team:Groningen/protocols/Motility_assay | Motility assay]]
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<br/>Sub1: Copmpetence medium (MC completed)
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<table border="1">
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<tr>
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<td>H20</td>
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<td>1,8 ml</td>
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<td></td>
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</tr>
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<tr>
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<td>10x MC (sub2)</td>
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<td>200 ul</td>
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<td> filter sterilize</td>
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</tr>
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<tr>
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<td>MgSO4</td>
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<td>6,7 ul</td>
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<td>autoclave sterilize</td>
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</tr>
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<tr>
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<td>Tryptophan 1%</td>
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<td>10 ul</td>
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<td>filter steralize, wrap in Al</td>
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</tr></table>
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<br/>Sub2: MC 10x
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<table border="1">
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<tr>
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<td>for</td>
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<td>100 ml</td>
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<td>10 ml</td>
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</tr>
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<tr>
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<td>K2H PO4</td>
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<td>14,036g</td>
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<td> 1,4036g</td>
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</tr>
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<tr>
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<td>KH2 PO$</td>
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<td>5,239g</td>
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<td>0,5239g</td>
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</tr>
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<tr>
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<td>Glucose</td>
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<td>20g</td>
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<td>2g</td>
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</tr>
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</table>
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Sub2: MC 10x
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Tri-Na Citrate 300mM (Sub3) 10ml 1ml
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Ferric NH4 citrate (Sub4) 1ml 0,1ml
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Casein Hydrolysate 1g 0,1g
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Potassium Glutamate 2g 0,2g
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Add 50ml H2O, Mix, add H2O till 100ml, Filter sterilize and freeze at -20 °C
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Sub3: Tri-Na Citrate 300mM
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Tri-Na Citrate 0,8823g
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H2O 10ml
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Wrap in aluminum
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Sub4: Ferric NH4 citrate
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Ferric NH4 citrate 0,22g
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H2O 10ml
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Wrap in aluminum
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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"