Team:Tokyo Metropolitan/Project/Fiber/Protocol
From 2010.igem.org
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=== Grow up a culture of A.xylinus in media recommend by JCM=== | === Grow up a culture of A.xylinus in media recommend by JCM=== | ||
- | <html><center><table><tr><td width="850"> | + | <html> |
- | <font size="3"><span style="text-decoration:underline">Material</font></span>< | + | <center><table><tr><td width="850" align="left"> |
- | + | <font size="3"><span style="text-decoration:underline">Material</font></span> | |
- | + | <ul><li>A.xylinus JCM strain 7664 | |
- | + | <li>500 ml of liquid Acetobacter media(Open Wet Ware recommended) | |
- | + | <ul><li>Glucose - 100g | |
- | + | <li>Yeast extract - 10g | |
- | + | <li>CaCO3 30g | |
- | (If you are making plates, use the same protocol but add | + | <li>Distilled water - 1000 ml |
+ | </ul></li></ul> | ||
+ | (If you are making plates, use the same protocol but add 7.5 g of agar.)<br><br> | ||
- | <font size="3"><span style="text-decoration:underline">Equipment</font></span>< | + | <font size="3"><span style="text-decoration:underline">Equipment</font></span> |
- | + | <ul><li>autoclave | |
- | + | <li>incubator | |
- | + | <li>scale | |
- | + | <li>bunsen burner | |
- | + | <li>flask(1l or500ml) | |
- | + | <li>plate | |
- | + | <li>spreader | |
- | + | <li>pipet | |
- | + | <li>pipet tip</ul><br> | |
<font size="3"><span style="text-decoration:underline">Procedure</font></span> | <font size="3"><span style="text-decoration:underline">Procedure</font></span> | ||
<ol><li>Prepare media as outlined (add the materials as above) | <ol><li>Prepare media as outlined (add the materials as above) | ||
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<font size="3"><span style="text-decoration:underline">Material</font></span><br> | <font size="3"><span style="text-decoration:underline">Material</font></span><br> | ||
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<font size="3"><span style="text-decoration:underline">Material</font></span><br> | <font size="3"><span style="text-decoration:underline">Material</font></span><br> | ||
E.coli K12 colony<br> | E.coli K12 colony<br> | ||
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<font size="3"><span style="text-decoration:underline">Material</font></span><br> | <font size="3"><span style="text-decoration:underline">Material</font></span><br> | ||
<font size="3"><span style="text-decoration:underline">Equipment</font></span><br> | <font size="3"><span style="text-decoration:underline">Equipment</font></span><br> | ||
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<font size="3"><span style="text-decoration:underline">Material</font></span><br> | <font size="3"><span style="text-decoration:underline">Material</font></span><br> | ||
QIAGEN(gel extraction kit)<br> | QIAGEN(gel extraction kit)<br> |
Revision as of 09:00, 29 August 2010
![](https://static.igem.org/mediawiki/2010/9/9f/Re-main.png)
E.coli Fiber Project Protocol
Contents |
Grow up a culture of A.xylinus in media
Grow up a culture of A.xylinus in media recommend by Open Wet Ware
Material
Equipment
Procedure
Note
|
Grow up a culture of A.xylinus in media recommend by JCM
Material
Equipment
Procedure
Note
|
Grow up a culture of E.coli
Material Equipment Procedure |
Direct PCR
Material E.coli K12 colony 10µM/l forward Primer 5µl 10µM/l reverse Primer 5µl 10×EX taq buffer 10µl 2.5mM each dNTP mixture 10µl EX taq polymerase 1µl milli-Q 71µl Equipment thermal cycler vortex mixer PCR-tubes pipet pipet tip Procedure
|
Electrophoreses PCR productions
Material Equipment Procedure |
DNA purification from agarose gel with QIAGEN
Material QIAGEN(gel extraction kit) -QG buffer 300µl -PE buffer700µl -EB buffer 50µl -tube for column Equipment centrifuge heating plate pipet pipet tip Procedure 1. cut gel of electrophoreses 2. add pieces of gel to tubes 3. take 300µl QG buffer into tubes and dissolve at 50°C 4. add a solution of QG buffer and gel to tubes for column 5. centrifuge 15000rpm/1min 6. throw “flow-thru” away and take 700µl PE buffer 7. centrifuge 15000rpm/1min 8. throw flow-thru away 9. centrifuge 15000rpm/1min 10. change tube for column 11. add 50µl of EB buffer (aim to center of tube) 12. centrifuge 15000rpm/1min |