Team:Tokyo Metropolitan/Project/Fiber/Protocol

From 2010.igem.org

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<html><font size="5">E.coli Fiber Project Protocol</font></html>
<html><font size="5">E.coli Fiber Project Protocol</font></html>
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==Grow up a culture of A.xylinus in media==
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===Grow up a culture of A.xylinus in media recommend by Open Wet Ware===
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==　Grow up a culture of A.xylinus in media==
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===Grow up a culture of A.xylinus in media recommend by JCM===
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===　Grow up a culture of A.xylinus in media recommend by Open Wet Ware===
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==Grow up a culture of E.coli==
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<html></html>
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==Direct PCR==
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===　Grow up a culture of A.xylinus in media recommend by JCM===
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==Electrophoreses PCR productions==
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<html></html>
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==DNA purification from agarose gel with QIAGEN==
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==　Grow up a culture of E.coli==
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<html></html>
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==　Direct PCR==
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<html></html>
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==　Electrophoreses PCR productions==
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<html></html>
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==　DNA purification from agarose gel with QIAGEN==
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<html>
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　<font size="3"><span style="text-decoration:underline">Material</font></span><br>
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　　QIAGEN(gel extraction kit)<br>
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　　-QG buffer 300µl<br>
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　　-PE buffer700µl<br>
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　　-EB buffer 50µl<br>
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　　-tube for column<br>
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　<font size="3"><span style="text-decoration:underline">Equipment</font></span><br>
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　　centrifuge<br>
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　　heating plate<br>
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　　pipet<br>
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　　pipet tip<br>
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　<font size="3"><span style="text-decoration:underline">Procedure</font></span><br>
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　1. cut gel of electrophoreses<br>
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　2. add pieces of gel to tubes<br>
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　3. take 300µl QG buffer into tubes and dissolve at 50°C<br>
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　4. add a solution of QG buffer and gel to tubes for column<br>
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　5. centrifuge 15000rpm/1min<br>
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　6. throw “flow-thru” away and take 700µl PE buffer<br>
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　7. centrifuge 15000rpm/1min<br>
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　8. throw flow-thru away <br>
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　9. centrifuge 15000rpm/1min<br>
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　10. change tube for column<br>
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　11. add 50µl of EB buffer (aim to center of tube)<br>
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　12. centrifuge 15000rpm/1min<br>
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</html>

Revision as of 18:53, 28 August 2010


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

　Grow up a culture of A.xylinus in media recommend by JCM

　Grow up a culture of E.coli

　Direct PCR

　Electrophoreses PCR productions

　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

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