Team:Gifu/Notebook
From 2014.igem.org
(Difference between revisions)
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<li>-5'intron M-His-RFP(without stop codon)-</li> | <li>-5'intron M-His-RFP(without stop codon)-</li> | ||
</ol> | </ol> | ||
- | <h3>Miniprep</h3> | + | <h3>Miniprep</h3> |
- | + | ||
<h3>Colony PCR</h3> | <h3>Colony PCR</h3> | ||
+ | <h3>Restriction Enzyme Digestion</h3> | ||
+ | <table> | ||
+ | <tr> | ||
+ | <th>sample</th><th>restriction enzymes</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>-His-RFP(with stop codon)-5'intron M-DT-Cm-</td><td>EcoRI,XbaI</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>-His-RFP(without stop codon)-5'intron M-DT-Cm-</td><td>EcoRI,XbaI</td> | ||
+ | </tr> | ||
+ | </table> | ||
<h3>Densitometry</h3> | <h3>Densitometry</h3> | ||
- | < | + | <table> |
- | <h3>Transformation</h3> | + | <tr> |
+ | <th>sample</th><th>concentration(ng/µL)</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>EcoRI-Lac-3'intron-RBS-SpeI</td><td>120.0</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>XbaI-His-RFP(with stop codon)-5'intron M-DT-Cm-EcoRI</td><td>187.8</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>XbaI-His-RFP(without stop codon)-5'intron M-DT-Cm-EcoRI</td><td>158.9</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <h3>Ligation&Transformation</h3> | ||
+ | <ol class="list"> | ||
+ | <li>EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(with stop codon)-5'intron M-DT-Cm-EcoRI</li> | ||
+ | <li>EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(without stop codon)-5'intron M-DT-Cm-EcoRI</li> | ||
+ | </ol> | ||
<h3>Analysis of DNA sequences</h3> | <h3>Analysis of DNA sequences</h3> | ||
<p align="right"><a href="#calender">back to calender</a></p> | <p align="right"><a href="#calender">back to calender</a></p> |
Revision as of 03:56, 3 September 2014
Calender
Click on a date to see our research activities on that day.
August | September | |||||||||||||||
Mon | Tue | Wed | Thr | Fri | Sat | Sun | Mon | Tue | Wed | Thr | Fri | Sat | Sun | |||
1 | 2 | 3 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | |||||||
4 | 5 | 6 | 7 | 8 | 9 | 10 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |||
11 | 12 | 13 | 14 | 15 | 16 | 17 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | |||
18 | 19 | 20 | 21 | 22 | 23 | 24 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | |||
25 | 26 | 27 | 28 | 29 | 30 | 31 | 29 | 30 | > |
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 |
August 12th
Colony PCR
- 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:[1]turning red
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[2]turning pink
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[3]turning white
- His-RFP(without stop codon)-Amp
Electrophoresis
- His-RFP(with stop codon)-DT
- His-RFP
- 5'intron M-DT
- 5'intron M
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[1]
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[2]
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[3]
- His-RFP(without stop codon)-5'intron-DT
- His-RFP(without stop codon)
Liquid Culture
- His-RFP(with stop codon)-DT
- 5'intron M-DT
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[1]
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[3]
- His-RFP(without stop codon)
August 13th
Miniprep
PCR
By use of KAPA HiFi DNA Polymerase- 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:[1]turning red
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[3]turning white
Electrophoresis
- His-RFP(with stop codon)-DT-Cm
- 5'intron M-DT-Cm
- DNA Ladder Markers DNA Ladder One(Broad Range)
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[1]turning red
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[3]turning white
Restriction Enzyme Digestion
sample | restriction enzymes |
---|---|
His-RFP(with stop codon)-DT-Cm | EcoRI,XbaI |
5'intron M-DT-Cm | EcoRI,XbaI |
Densitometry
sample | concentration(ng/µL) |
---|---|
EcoRI-Lac-RBS-SpeI | 65.0 |
XbaI-His-RFP(with stop codon)-DT-Cm-EcoRI | 188.8 |
EcoRI-His-RFP(with stop codon)-SpeI | 160.0 |
EcoRI-His-RFP(without stop codon)-SpeI | 160.0 |
XbaI-5'intron M-DT-Cm-EcoRI | 167.8 |
Ligation
- EcoRI-Lac-RBS-SpeI + XbaI-His-RFP(with stop codon)-DT-Cm-EcoRI
- EcoRI-His-RFP(with stop codon)-SpeI + XbaI-5'intron M-DT-Cm-EcoRI
- EcoRI-His-RFP(without stop codon)-SpeI + XbaI-5'intron M-DT-Cm-EcoRI
Transformation
- -Lac-RBS-His-RFP(with stop codon)-DT-Cm-
- -His-RFP(with stop codon)-5'intron M-DT-Cm-
- -His-RFP(without stop codon)-5'intron M-DT-Cm-
August 16th
Liquid Culture
August 18th
Plasmid Extraction
- -Lac-RBS-His-RFP(with stop codon)-DT-Cm-
- -His-RFP(with stop codon)-5'intron M-DT-Cm-
- -His-RFP(without stop codon)-5'intron M-DT-Cm-
Plasmid Purification
- -5'intron M-His-RFP(with stop codon)-
- -5'intron M-His-RFP(without stop codon)-
Miniprep
Colony PCR
Restriction Enzyme Digestion
sample | restriction enzymes |
---|---|
-His-RFP(with stop codon)-5'intron M-DT-Cm- | EcoRI,XbaI |
-His-RFP(without stop codon)-5'intron M-DT-Cm- | EcoRI,XbaI |
Densitometry
sample | concentration(ng/µL) |
---|---|
EcoRI-Lac-3'intron-RBS-SpeI | 120.0 |
XbaI-His-RFP(with stop codon)-5'intron M-DT-Cm-EcoRI | 187.8 |
XbaI-His-RFP(without stop codon)-5'intron M-DT-Cm-EcoRI | 158.9 |
Ligation&Transformation
- EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(with stop codon)-5'intron M-DT-Cm-EcoRI
- EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(without stop codon)-5'intron M-DT-Cm-EcoRI
Analysis of DNA sequences
August 19th
Colony PCR
Electrophoresis
Shaking Culture
August 20th
Colony PCR
Same as August 19th Colony PCR.