Team:KAIST Korea/Notebook Protocol
From 2012.igem.org
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<h3>Materials</h3> | <h3>Materials</h3> | ||
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<h4>Pellet Cells</h4> | <h4>Pellet Cells</h4> | ||
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<li>Remove the supernatant.</li> | <li>Remove the supernatant.</li> | ||
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<h4>Lyse Cells</h4> | <h4>Lyse Cells</h4> | ||
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<li>Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.</li> | <li>Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.</li> | ||
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<li>Remove the supernatant.</li> | <li>Remove the supernatant.</li> | ||
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<h4>Protein Precipitation</h4> | <h4>Protein Precipitation</h4> | ||
+ | </br> | ||
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<li>Add 200ul of Protein Precipitation Solution to the RNase-treated cell lysate. Vortex vigorously at high speed for 20seconds to mix the Protein Precipitation Solution tieh the cell lysate.</li> | <li>Add 200ul of Protein Precipitation Solution to the RNase-treated cell lysate. Vortex vigorously at high speed for 20seconds to mix the Protein Precipitation Solution tieh the cell lysate.</li> | ||
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<li>Add 50~100 μl TE buffer and rehydrate the DNA by incubating at 65℃ for 20min. Periodically mix the solution by gently tapping the tube. Alternatively, rehydrate the DNA by incubationg the solution overnight at room temperature or at 4℃.</li> | <li>Add 50~100 μl TE buffer and rehydrate the DNA by incubating at 65℃ for 20min. Periodically mix the solution by gently tapping the tube. Alternatively, rehydrate the DNA by incubationg the solution overnight at room temperature or at 4℃.</li> | ||
</ol> | </ol> | ||
+ | </br> | ||
<img id="figure" alt="materials" src="https://static.igem.org/mediawiki/2012/d/df/KAIST_Protocol2_table2.png"></img> | <img id="figure" alt="materials" src="https://static.igem.org/mediawiki/2012/d/df/KAIST_Protocol2_table2.png"></img> | ||
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<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | ||
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<span id="starter">P</span>rotocol 3 content<br /><br /><br /><br /><br /><br /> | <span id="starter">P</span>rotocol 3 content<br /><br /><br /><br /><br /><br /> | ||
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<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | ||
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<span id="starter">P</span>rotocol 4 content<br /><br /><br /><br /><br /><br /> | <span id="starter">P</span>rotocol 4 content<br /><br /><br /><br /><br /><br /> | ||
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<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | ||
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<span id="starter">P</span>rotocol 5 content<br /><br /><br /><br /><br /><br /> | <span id="starter">P</span>rotocol 5 content<br /><br /><br /><br /><br /><br /> | ||
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<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | ||
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<span id="starter">P</span>rotocol 6 content<br /><br /><br /><br /><br /><br /> | <span id="starter">P</span>rotocol 6 content<br /><br /><br /><br /><br /><br /> | ||
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<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/> | ||
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<span id="starter">P</span>rotocol 7 content 아졸려 언제와 점심먹고 왔는데 아무도 없어<br /><br /><br /><br /><br /><br /> | <span id="starter">P</span>rotocol 7 content 아졸려 언제와 점심먹고 왔는데 아무도 없어<br /><br /><br /><br /><br /><br /> | ||
</div> | </div> |
Revision as of 08:44, 22 August 2012
2012 KAIST Korea
Mail : kaist.igem.2012@gmail.com
Twitter : twitter.com/KAIST_iGEM_2012
Facebook : www.facebook.com/KAISTiGEM2012
image description
Lab Protocols
Please enter sub title1. LB agar plate
Materials
Procedure
- Add 25g LB broth and 15g agar into 1L DDW.
- Autoclave
2. Genomic DNA Purification
Materials
Procedure
Pellet Cells
- Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.
- Centrifuge at 13000 rpm for 2.5minutes to pellet the cells.
- Remove the supernatant.
Lyse Cells
- Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.
- Centrifuge at 13000 rpm for 2.5minutes to pellet the cells.
- Remove the supernatant.
Protein Precipitation
- Add 200ul of Protein Precipitation Solution to the RNase-treated cell lysate. Vortex vigorously at high speed for 20seconds to mix the Protein Precipitation Solution tieh the cell lysate.
- Incubate the sample on ice for 5minutes.
- Centrifuge at 13,000rpm 3minutes.
- Transfer the supernatant containing the DNA to a clean 1.5ml microcentrifuge tube. Note: Some supernatant may remain in the original tube containing the protein pellet. Leave this residual liquid in the tube to avoid contaminating the DNA solution with the precipitated protein.
- Add 600ul isopropanol.
- Gently mix by inversion until the thread-like strands of DNA form a visible mass.
- Centrifuge at 13000rpm for 2minutes.
- Carefully pour off the supernatant.
- Add 600ul of room temperature 70% ethanol and gently invert the tube 5 times to wash the DNA pellet.
- Centrifuge at 13000rpm for 2minutes. Carefully aspirate the ethanol.
- Drain the tube on clean absorbent paper and allow the pellet to air-dry for 10-14minutes.
- Add 50~100 μl TE buffer and rehydrate the DNA by incubating at 65℃ for 20min. Periodically mix the solution by gently tapping the tube. Alternatively, rehydrate the DNA by incubationg the solution overnight at room temperature or at 4℃.
Protocol 3 title
Protocol 3 content
Protocol 4 title
Protocol 4 content
Protocol 5 title
Protocol 5 content
Protocol 6 title
Protocol 6 content
Protocol 7 title
Protocol 7 content 아졸려 언제와 점심먹고 왔는데 아무도 없어