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Team:Gifu/Notebook
From 2014.igem.org
(Difference between revisions)
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</tbody> | </tbody> | ||
</table> | </table> | ||
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<h2><a name="0812"></a>August 12th</h2> | <h2><a name="0812"></a>August 12th</h2> | ||
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<li>His-RFP(without stop codon)</li> | <li>His-RFP(without stop codon)</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0813"></a>August 13th</h2> | <h2><a name="0813"></a>August 13th</h2> | ||
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<li>-His-RFP(without stop codon)-5'intron M-DT-Cm-</li> | <li>-His-RFP(without stop codon)-5'intron M-DT-Cm-</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0816"></a>August 16th</h2> | <h2><a name="0816"></a>August 16th</h2> | ||
<h3>Liquid Culture </h3> | <h3>Liquid Culture </h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0818"></a>August 18th</h2> | <h2><a name="0818"></a>August 18th</h2> | ||
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</ol> | </ol> | ||
<h3>Polyethylene glycol Precipitation </h3> | <h3>Polyethylene glycol Precipitation </h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0819"></a>August 19th</h2> | <h2><a name="0819"></a>August 19th</h2> | ||
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</ol> | </ol> | ||
<h3>Shaking Culture</h3> | <h3>Shaking Culture</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0820"></a>August 20th</h2> | <h2><a name="0820"></a>August 20th</h2> | ||
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<li>-His-RFP(without stop codon)-5'intron M-DT-Cm-</li> | <li>-His-RFP(without stop codon)-5'intron M-DT-Cm-</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0821"></a>August 21th</h2> | <h2><a name="0821"></a>August 21th</h2> | ||
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<h3>Protein Extraction </h3> | <h3>Protein Extraction </h3> | ||
<h3>SDS-PAGE</h3> | <h3>SDS-PAGE</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0822"></a>August 22th</h2> | <h2><a name="0822"></a>August 22th</h2> | ||
<h3>Prorein Purification by use of Ni-NTA</h3> | <h3>Prorein Purification by use of Ni-NTA</h3> | ||
<h3>SDS-PAGE</h3> | <h3>SDS-PAGE</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0823"></a>August 23th</h2> | <h2><a name="0823"></a>August 23th</h2> | ||
<h3>Prorein Purification by use of Ni-NTA</h3> | <h3>Prorein Purification by use of Ni-NTA</h3> | ||
<h3>SDS-PAGE</h3> | <h3>SDS-PAGE</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0824"></a>August 24th</h2> | <h2><a name="0824"></a>August 24th</h2> | ||
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<li>EcoRI-Lac-3'intron-RBS-PstI</li> | <li>EcoRI-Lac-3'intron-RBS-PstI</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0825"></a>August 25th</h2> | <h2><a name="0825"></a>August 25th</h2> | ||
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<li>Lac-3'intron-RBS-His-RFP-5'intron M-DT-Cm</li> | <li>Lac-3'intron-RBS-His-RFP-5'intron M-DT-Cm</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0826"></a>August 26th</h2> | <h2><a name="0826"></a>August 26th</h2> | ||
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<li>EcoRI-TTHA0175-SpeI + EcoRI-5'intron M-DT-Cm-XbaI</li> | <li>EcoRI-TTHA0175-SpeI + EcoRI-5'intron M-DT-Cm-XbaI</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0827"></a>August 27th</h2> | <h2><a name="0827"></a>August 27th</h2> | ||
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</ol> | </ol> | ||
<h3>Enrichment Culture </h3> | <h3>Enrichment Culture </h3> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0828"></a>August 28th</h2> | <h2><a name="0828"></a>August 28th</h2> | ||
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<li>-5'intron M-Cm-</li> | <li>-5'intron M-Cm-</li> | ||
</ol> | </ol> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0829"></a>August 29th</h2> | <h2><a name="0829"></a>August 29th</h2> | ||
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<li>EcoRI-TTHA0175-5'intron M-DT-Cm-XbaI</li> | <li>EcoRI-TTHA0175-5'intron M-DT-Cm-XbaI</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0830"></a>August 30th</h2> | <h2><a name="0830"></a>August 30th</h2> | ||
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<li>Plate4 3G</li> | <li>Plate4 3G</li> | ||
</ol> | </ol> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0831"></a>August 31th</h2> | <h2><a name="0831"></a>August 31th</h2> | ||
<h3>Colony PCR</h3> | <h3>Colony PCR</h3> | ||
<p> Because result of Colony PCR on 8/30 is not good, We chose 5 colonies and put them on PCR.</p> | <p> Because result of Colony PCR on 8/30 is not good, We chose 5 colonies and put them on PCR.</p> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0901"></a>September 1st</h2> | <h2><a name="0901"></a>September 1st</h2> | ||
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</ol> | </ol> | ||
<h3>Cell body Crush&SDS-PAGE(gel 10%)</h3> | <h3>Cell body Crush&SDS-PAGE(gel 10%)</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0903"></a>September 3rd</h2> | <h2><a name="0903"></a>September 3rd</h2> | ||
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<li>-Lac-3'intron-RBS-RFP(with stop codon)-5'intron M-DT-Cm- 5 mL</li> | <li>-Lac-3'intron-RBS-RFP(with stop codon)-5'intron M-DT-Cm- 5 mL</li> | ||
</ol> | </ol> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0904"></a>September 4th</h2> | <h2><a name="0904"></a>September 4th</h2> | ||
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<li>RFP(with stop codon)</li> | <li>RFP(with stop codon)</li> | ||
</ol> | </ol> | ||
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<h2><a name="0905"></a>September 5th</h2> | <h2><a name="0905"></a>September 5th</h2> | ||
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<li>RFP(with stop codon) supernatant3</li> | <li>RFP(with stop codon) supernatant3</li> | ||
</ol> | </ol> | ||
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<h2><a name="0908"></a>September 8th</h2> | <h2><a name="0908"></a>September 8th</h2> | ||
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<h3></h3> | <h3></h3> | ||
<h3></h3> | <h3></h3> | ||
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<h2><a name="0909"></a>September 9th</h2> | <h2><a name="0909"></a>September 9th</h2> | ||
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<h3></h3> | <h3></h3> | ||
<h3></h3> | <h3></h3> | ||
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<h2><a name="0910"></a>September 10th</h2> | <h2><a name="0910"></a>September 10th</h2> | ||
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<li>guanidine deposition</li> | <li>guanidine deposition</li> | ||
</ol> | </ol> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0911"></a>September 11th</h2> | <h2><a name="0911"></a>September 11th</h2> | ||
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<li>TTHA0715 guanidine deposition</li> | <li>TTHA0715 guanidine deposition</li> | ||
</ol> | </ol> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0912"></a>September 12th</h2> | <h2><a name="0912"></a>September 12th</h2> | ||
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<li>-Lac-RBS-GFP-DT-Cm-</li> | <li>-Lac-RBS-GFP-DT-Cm-</li> | ||
</ol> | </ol> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0913"></a>September 13th</h2> | <h2><a name="0913"></a>September 13th</h2> | ||
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<h3>cDNA synthesis</h3> | <h3>cDNA synthesis</h3> | ||
<h3>PCR for confirming existence of circular RNA</h3> | <h3>PCR for confirming existence of circular RNA</h3> | ||
| - | <noscript> | + | <br><noscript> |
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0914"></a>September 14th</h2> | <h2><a name="0914"></a>September 14th</h2> | ||
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</ol> | </ol> | ||
<h3>Ethanol Precipitation</h3> | <h3>Ethanol Precipitation</h3> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0915"></a>September 15th</h2> | <h2><a name="0915"></a>September 15th</h2> | ||
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<h3>reverse transcription PCR</h3> | <h3>reverse transcription PCR</h3> | ||
<h3>PCR</h3> | <h3>PCR</h3> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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<h2><a name="0916"></a>September 16th</h2> | <h2><a name="0916"></a>September 16th</h2> | ||
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<h3>PCR for confirming existence of circular RNA</h3> | <h3>PCR for confirming existence of circular RNA</h3> | ||
<h3></h3> | <h3></h3> | ||
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<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> | ||
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Revision as of 00:54, 25 September 2014
Calender
Click on a date to see our research activities on that day.
| August | ||||||
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| Mon | Tue | Wed | Thr | Fri | Sat | Sun |
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| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | 26 | 27 | 28 | 29 | 30 | 31 |
| September | ||||||
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| Mon | Tue | Wed | Thr | Fri | Sat | Sun |
| 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 8 | 9 | 10 | 11 | 12 | 13 | 14 |
| 15 | 16 | 17 | 18 | 19 | 20 | 21 |
| 22 | 23 | 24 | 25 | 26 | 27 | 28 |
| 29 | 30 | |||||
| October | ||||||
|---|---|---|---|---|---|---|
| Mon | Tue | Wed | Thr | Fri | Sat | Sun |
| 1 | 2 | 3 | 4 | 5 | ||
| 6 | 7 | 8 | 9 | 10 | 11 | 12 |
| 13 | 14 | 15 | 16 | 17 | 18 | 19 |
| 20 | 21 | 22 | 23 | 24 | 25 | 26 |
| 27 | 28 | 29 | 30 | 31 | ||
Glossary
- His
- alphabet
- GFP
- Green Fluorescence Protein
- RFP
- Red Fluorescent ProteinThis is BBa_E1010.
- 5'-intron
- 5'-intron M
- 3'-intron
- Lac
- LacI regulated promoter.This is BBa_R0010.
- Amp
- Ampicillin resistance gene
- Cm
- Chloramphenicol resistance gene.This is pSB1C3.
- RBS
- Ribosome Binding Sites.This is BBa_B0034.
- DT
- Double Terminator consisting of BBa_B0010 and BBa_B0012. This is BBa_B0015.
- EcoRI
- Restriction site that is cut by EcoRI or restriction enzyme
- XbaI
- Restriction site that is cut by XbaI or restriction enzyme
- SpeI
- Restriction site that is cut by SpeI or restriction enzyme
- PstI
- Restriction site that is cut by PstI or restriction enzyme
- TTHA0715
- Cold shock protein with amino acid sequence cut by thrombin
- TTHA0715 guanidine deposition
- SmtA
- circular RNA
Prologue -before August-
Parts amplification with PCR
- Lac
- RBS
- RFP
- DT
- Cm (pSB1C3)
- pSB1A3
- pSB1K3
T4 phage Multiplication with E.coli
DNA Extraction from T4 phage
August
August 11th
Restriction Enzyme Digestion
| sample | restriction enzymes |
|---|---|
| His-RFP(with stop codon) | SpeI,EcoRI | 5'-intron M | SpeI,EcoRI |
| Lac-3'intron-RBS | SpeI,EcoRI |
| His-RFP(without stop codon) | XbaI,EcoRI |
| RBS-Amp | SpeI,EcoRI |
Ligation
- EcoRI-His-RFP(with stop codon)-SpeI + XbaI-DT-Cm-EcoRI
- EcoRI-5'intron M-SpeI + XbaI-DT-Cm-EcoRI
- EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(without stop codon)-5'intron-DT-Cm-EcoRI
- EcoRI-His-RFP(without stop codon)-SpeI + SpeI-Amp-EcoRI
Transformation
- -His-RFP(with stop codon)-DT-Cm-
- -5'intron M-DT-Cm-
- -Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm-
- -His-RFP(without stop codon)-Amp-
Densitometry
| sample | concentration(ng/µL) |
|---|---|
| His-RFP(with stop codon) EcoRI-SpeI | 162.6 |
| 5'intron M EcoRI-SpeI | 45.4 |
| Lac-3'intron-RBS EcoRI-SpeI | 120.9 |
| His-RFP(without stop codon) EcoRI-SpeI | 160.0 |
| His-RFP(without stop codon)-5'intron-DT-Cm XbaI-EcoRI | 174.0 |
| Amp EcoRI-SpeI | 101.8 |
