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| - | {{Kyoto_Foreground}} | + | {{Kyoto_Foreground|active_page=notebook}} |
| | + | {{Kyoto_Background}} |
| | {{Kyoto_WikiDesign}} | | {{Kyoto_WikiDesign}} |
| | | | |
| - | <!-- #main -->
| + | = Notebook = |
| - | <div id="main">
| + | |
| | | | |
| - | = Lab Note =
| + | See our daily progress with the project ! |
| | | | |
| | <html> | | <html> |
| | <style> | | <style> |
| - | #index {
| + | a.note_link:visited { |
| - | margin:10px 0;
| + | color: #ffffff; |
| - | padding:0;
| + | } |
| - | width:700px;
| + | a.note_link { |
| - | height:30px;
| + | |
| - | overflow:hidden;
| + | |
| - | }
| + | |
| - | #index li { list-style:none; width:68px; float:left; } | + | |
| - | #index li a {
| + | |
| - | display:block;
| + | |
| - | line-height:30px;
| + | |
| - | text-align:center;
| + | |
| - | } | + | |
| - | | + | |
| - | #index li a {
| + | |
| | display: block; | | display: block; |
| - | line-height:30px; | + | margin: 10px; |
| | + | width: 375px; |
| | + | height: 50px; |
| | + | line-height:50px; |
| | + | text-align: center; |
| | + | background-color: #0000ff; |
| | + | color: #ffffff; |
| | + | font-size: 15px; |
| | } | | } |
| | </style> | | </style> |
| - | <ul id="index">
| + | |
| - | <li><a href=" #lab-week1">Week1</ a></ li> | + | <a href="https://2011.igem.org/Team:Kyoto/Lab Work" class="note_link">Lab Work</span></a> |
| - | <li><a href="#lab-week2">Week2</a></li>
| + | <a href="https://2011.igem.org/Team:Kyoto/Diary" class="note_link">Diary</span></a> |
| - | <li><a href="#lab-week3">Week3</a></li>
| + | |
| - | <li><a href="#lab-week4">Week4</a></li>
| + | |
| - | <li><a href="#lab-week5">Week5</a></li>
| + | |
| - | <li><a href=" #lab-week6">Week6</ a></ li> | + | |
| - | <li><a href="#lab-week7">Week7</a></li>
| + | |
| - | <li><a href="#lab-week8">Week8</a></li>
| + | |
| - | <li><a href="#lab-week9">Week9</a></li>
| + | |
| - | <li><a href="#protocol">Protocol</a></li>
| + | |
| - | </ul>
| + | |
| | </html> | | </html> |
| - |
| |
| - | <html><a name="lab-week1"></a></html>
| |
| - | == Week1: Monday 1st - Sunday 7th August ==
| |
| - | '''Monday'''<br/>
| |
| - | 行った実験名：
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| - |
| |
| - | 使った試薬名、容量：
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| - |
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| - | 用いた機械：
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| - |
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| - | 行った人：
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| - |
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
| |
| - | <html><a name="lab-week2"></a></html>
| |
| - |
| |
| - | == Week2: Monday 8th - Sunday 14th August ==
| |
| - | '''Monday'''<br/>
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
| |
| - | <html><a name="lab-week3"></a></html>
| |
| - | == Week3: Monday 15th - Sunday 21th August ==
| |
| - | '''Monday'''<br/>
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| - |
| |
| - | '''Tuesday'''<br/>
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| - |
| |
| - | '''Wednesday'''<br/>
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| - |
| |
| - | '''Thursday'''<br/>
| |
| - | Nutritional Signal(Sugiura,Shimosaka & Okumura)<br/>
| |
| - | PCR Amplification of glnL and glnG from gDNA of E.coli<br/>
| |
| - |
| |
| - | --Primers--<br/>
| |
| - | glnL<br/>
| |
| - | Left primer: tctagaggagactgctttatggcaac<br/>
| |
| - | Right primer: actagtaggaactatcgtcatcgactac<br/>
| |
| - | glnG<br/>
| |
| - | Left primer: tctagaggtgacgtttatgcaacga<br/>
| |
| - | Right primer: actagtacacacaagctgtgaatcactc<br/>
| |
| - | annealing temperature was 55 degrees.<br/>
| |
| - | [[File:Kyoto-Gel0818.jpg]]<br/>
| |
| - | lane 1,2 &7,8: DNA ladder(λDNA digested Hindlll,100bp), lane 3,4: glnG1,glnG2, lane 5,6: glnL1,glnL2<br/>
| |
| - |
| |
| - | After purification, the concentration of DNA are<br/>
| |
| - | glnG1: 127.8ng/ul<br/>
| |
| - | glnG2: 118.1ng/ul<br/>
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| - | glnL1: 137.4ng/ul<br/>
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| - | glnL2: 124.2ng/ul<br/>
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| - |
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| - | '''Friday'''<br/>
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| - | Nutritional Signal(Shimosaka)<br/>
| |
| - | Restriction of glnG1 and glnG2<br/>
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| - | Cut them with Xbal and Spel for 2 hours at 37 degrees.<br/>
| |
| - | Then, gel extraction of digested.<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
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| - | <html><a name="lab-week4"></a></html>
| |
| - |
| |
| - | == Week4: Monday 22th - Sunday 28th August ==
| |
| - | '''Monday'''<br/>
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
| |
| - | Luminescence(Kusaba, Terada, Hara):ハエの走行性実験① ♂、紫外線×2回、緑×2回 ♀、紫外線×2回、緑×2回
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| - |
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| - | '''Friday'''<br/>
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| - | Luminescence(Kusaba):ハエの走行性実験① ♂、赤外線×2回 ♀、赤外線×2回
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| - |
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| - | Nutritional Signal(Hashiya):
| |
| - | Transformation of bellow parts.<br/>
| |
| - | 4-17M:BBa_K325909(lux operon)<br/>
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| - | 1-12M:BBa_E0240<br/>
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| - | 2-17F:BBa_120260(low copy vector)<br/>
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| - |
| |
| - | PCR amplification of
| |
| - |
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| - | '''Saturday'''<br/>
| |
| - | Luminescence(Kusaba):ハエの走行性実験① ♂、赤×2回、青×2回 ♀、赤×2回、青×2回
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| - |
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| - | '''Sunday'''<br/>
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| - |
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| - | <html><a name="lab-week5"></a></html>
| |
| - |
| |
| - | == Week5: Monday 29th August - Sunday 4th September ==
| |
| - | '''Monday'''<br/>
| |
| - |
| |
| - | '''Tuesday'''<br/>
| |
| - | Nutritional Signal(Hashiya)<br/>
| |
| - | ・PCR amplification of glnL and glnG from PCR products,glnL1 and glnG1.<br/>
| |
| - | --Primers--<br/>
| |
| - | glnL<br/>
| |
| - | Left primer: ggaattcgcggccgcttctagaggagactgctttatggcaac<br/>
| |
| - | Right primer: ggactagtaggaactatcgtcatcgactac<br/>
| |
| - | glnG<br/>
| |
| - | Left primer: ggaattcgcggccgcttctagaggtgacgtttatgcaacga<br/>
| |
| - | Right primer: ggactagtacacacaagctgtgaatcactc<br/>
| |
| - | annealing temperature was 55 degrees.<br/>
| |
| - |
| |
| - | ・PCR amplification of glnL+G and rpoN from gDNA of E.coli.<br/>
| |
| - | --Primers--<br/>
| |
| - | glnL+G<br/>
| |
| - | Left primer: ggaattcgcggccgcttctagaggagactgctttatggcaac<br/>
| |
| - | Right primer: ggactagtacacacaagctgtgaatcactc<br/>
| |
| - | rpoN<br/>
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| - | Left primer: ggaattcgcggccgcttctagaggttctgaacatgaagcaa<br/>
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| - | Right primer: ggactagtatccttatcggttgggtca<br/>
| |
| - | annealing temperature was 56 degrees.<br/>
| |
| - |
| |
| - | [[File:Kyoto-Gel08300.jpg]]<br/>
| |
| - | lane1: 100bp DNA ladder, lane2:glnL, lane3:glnG, lane4:glnG+L, lane5:rpoN from gDNA, lane6:rpoN from ASKA clone<br/>
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| - |
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| - | After purification, the concentration of DNA are<br/>
| |
| - | glnL: 122.3 ng/ul<br/>
| |
| - | glnG: 64.7 ng/ul<br/>
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| - | glnL+G: 106.7 ng/ul<br/>
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| - | rpoN from gDNA: 111.4 ng/ul<br/>
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| - |
| |
| - | '''Wednesday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Screening PCR of σ54 promoter + pSB1A3<br/>
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| - | [[File:Kyoto-Gel08301.jpg]]<br/>
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| - | We cultured σ54 promoter5<br/>
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| - |
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| - | '''Friday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Restriction of σ54 promoter5,glnL, glnG, glnL+G and rpoN<br/>
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| - | Cut them with EcoRl and Spel
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| - | After purification, the concentration of DNA were<br/>
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| - | σ54 promoter5: 23.6 ng/ul<br/>
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| - | glnL: 28.1 ng/ul<br/>
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| - | glnG: 26.3 ng/ul<br/>
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| - | glnL+G: 15.3 ng/ul<br/>
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| - | rpoN: 20.8 ng/ul<br/>
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| - |
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| - | ・Ligation reaction<br/>
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| - | Ligated glnL, glnG and rpoN to pSB1K3.<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Screening PCR of glnL, glnG, glnL+G and rpoN<br/>
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| - | glnL
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| - | [[File:Kyoto-Gel09020.jpg]]<br/>
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| - | glnG
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| - | [[File:Kyoto-Gel09021.jpg]]<br/>
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| - | glnL+G & rpoN
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| - | [[File:Kyoto-Gel09022.jpg]]<br/>
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| - | We cultured glnL5, glnG4<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Mini prep of glnL5 and glnG4<br/>
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| - | glnL5: 43.9 ng/ul<br/>
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| - | glnG4: 38.1 ng/ul<br/>
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| - |
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| - | ・Screening PCR of rpoN
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| - | [[File:Kyoto-Gel09020.jpg]]<br/>
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| - |
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| - | Predation(Hashiya)<br/>
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| - | ・PCR amplification of glmS<br/>
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| - | --Primers--<br/>
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| - | left primer:ggaattcgcggccgcttctagagcaggttgaccgacaacgata<br/>
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| - | right primer:ggactagtacgcagggcatccatttat<br/>
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| - | [[File:Kyoto-Gel09030.jpg]]<br/>
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| - | lane1: 100bp DNA ladder, lane2: glmS from gDNA, lane3: glmS from ASKA clone<br/>
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| - |
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| - | ・TA cloning of glmS<br/>
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| - | Ligated glmS from gDNA to pTA vector.<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Screening PCR of rpoN<br/>
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| - | [[File:Kyoto-Gel09020.jpg]]<br/>
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| - |
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| - | ・Transformation of bellow parts<br/>
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| - | 1-23L: BBa_B0015 (double terminator)<br/>
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| - | 1-18E: BBa_J23101 (constitutive promoter)<br/>
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| - | 1-18C: BBa_J23100 (constitutive promoter)<br/>
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| - |
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| - | <html><a name="lab-week6"></a></html>
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| - |
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| - | == Week6: Monday 5th September - Sunday 11th September ==
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| - | '''Monday'''<br/>
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| - | Nutritional Signal(Hashiya)<br/>
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| - | ・Screening PCR of glnL+G+double terminator<br/>
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| - |
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| - |
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| - | '''Tuesday'''<br/>
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| - | Luminescence(Kusaba, Hara):ハエの走行性実験②（②は改良版） ♂、緑×2回 ♀、青×1回
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| - |
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| - | '''Wednesday'''<br/>
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| - | Luminescence(Hara):ハエの走行性実験② ♂、青×2回 ♀、緑×2回、青×1回
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| - |
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| - | '''Thursday'''<br/>
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| - | Luminescence(Hara):ハエの走行性実験② ♂、紫外線×3回 ♀、紫外線×3回
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - | Luminescence(Kusaba, Hara):ハエの走行性実験② ♂、青×2回、赤×2回 ♀、青×2回、赤×2回
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| - |
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| - | '''Sunday'''<br/>
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| - | Luminescence(Kusaba, Hara):ハエの走行性実験② ♂、紫外線×2回、
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| - | <html><a name="lab-week7"></a></html>
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| - |
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| - | == Week7: Monday 12th September - Sunday 18th September ==
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| - | '''Monday'''<br/>
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| - |
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| - | Luminescence:大腸菌の形質転換(Hashiya) ハエの走行性実験②(Kusaba, Hara)
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | Luminescence:大腸菌はじめて光る。しかし光量は少ない。
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
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| - | <html><a name="lab-week8"></a></html>
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| - |
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| - | == Week8: Monday 19th September - Sunday 25th September ==
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| - | '''Monday'''<br/>
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
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| - | <html><a name="lab-week9"></a></html>
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| - | == Week9: Monday 26th September - Sunday 2nd October ==
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| - | '''Monday'''<br/>
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| - |
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| - | '''Tuesday'''<br/>
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| - |
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| - | '''Wednesday'''<br/>
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| - |
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| - | '''Thursday'''<br/>
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| - |
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| - | '''Friday'''<br/>
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| - |
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| - | '''Saturday'''<br/>
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| - |
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| - | '''Sunday'''<br/>
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| - |
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| - | <html><a name="protocol"></a></html>
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| - | == Protocol ==
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| - |
| |
| - | === Medium for drosophila ===
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| - | ::{| class="wikitable"
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| - | |+ [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila]
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| - | ! width="300" |Materials
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| - | ! width="300" |Methods
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| - | |-
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| - | |
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| - | * water : 500mL
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| - | * dry yeast : 20g
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| - | * corn flour : 45g
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| - | * glucose : 50g
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| - | * agarose : 3.5~5g
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| - | * propionic acid : 1.5mL
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| - | * 10% p-hydroxybenzoate in 70% Eternol : 5g
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| - | |
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| - | # Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it.
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| - | # Stir corn flour and glucose with the remaining water.
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| - | # Stir 1 and 2, then autoclave it again.
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| - | # after autoclave, add propionic acid and 10% p-hydroxybenzoate in 70% Eternol into it.■
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| - | |}
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| - |
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| - |
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| - |
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| - |
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| - |
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| - |
| |
| - | </div>
| |
| - | <!-- /#main -->
| |
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