@@ Line 5: / Line 5: @@
 <div id="main">
-= Lab Note =
+= Notebook =
+See our daily progress with the project !
 <html>
 <style>
-#index {
+.note_link {
-    margin:10px 0;
-    padding:0;
-    width:700px;
-    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;
-  line-height:30px;
+ margin: 10px;
+ width: 375px;
+ height: 50px;
+  line-height:50px;
+ text-align: center;
+ background-color: #0000ff;
+ color: #ffffff;
+ font-size: 15px;
 }
 </style>
-<ul id="index">
-<li><a href="#lab-week1">Week1</a></li>
+<a href="https://2011.igem.org/Team:Kyoto/Lab Work" style="float: left"><span class="note_link">Lab Work</span></a>
-<li><a href="#lab-week2">Week2</a></li>
+<a href="https://2011.igem.org/Team:Kyoto/Diary" style="float:right"><span 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><a name="lab-week1"></a></html>
-== Week1: Monday 1st - Sunday 7th August ==
-'''Monday'''<br/>
-行った実験名：
-使った試薬名、容量：
-用いた機械：
-行った人：
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''Thursday'''<br/>
-'''Friday'''<br/>
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="lab-week2"></a></html>
-== Week2: Monday 8th - Sunday 14th August ==
-'''Monday'''<br/>
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''Thursday'''<br/>
-'''Friday'''<br/>
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="lab-week3"></a></html>
-== Week3: Monday 15th - Sunday 21th August ==
-'''Monday'''<br/>
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''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/>
-glnL1: 137.4ng/ul<br/>
-glnL2: 124.2ng/ul<br/>
-'''Friday'''<br/>
-Nutritional Signal(Shimosaka)<br/>
-Restriction of glnG1 and glnG2<br/>
-Cut them with Xbal and Spel for 2 hours at 37 degrees.<br/>
-Then, gel extraction of digested.<br/>
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="lab-week4"></a></html>
-== Week4: Monday 22th - Sunday 28th August ==
-'''Monday'''<br/>
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''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
-'''Friday'''<br/>
-Luminescence(Kusaba):ハエの走行性実験①　♂、赤外線×2回　♀、赤外線×2回
-Nutritional Signal(Hashiya):
-Transformation of bellow parts.<br/>
--17M:BBa_K325909(lux operon)<br/>
--12M:BBa_E0240<br/>
--17F:BBa_120260(low copy vector)<br/>
-PCR amplification of
-'''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
-'''Sunday'''<br/>
-<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/>
-　Left primer: ggaattcgcggccgcttctagaggttctgaacatgaagcaa<br/>
-　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/>
-　After purification, the concentration of DNA are<br/>
-　glnL: 122.3 ng/ul<br/>
-　glnG: 64.7 ng/ul<br/>
-　glnL+G: 106.7 ng/ul<br/>
-　rpoN from gDNA: 111.4 ng/ul<br/>
-'''Wednesday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Screening PCR of σ54 promoter + pSB1A3<br/>
-　[[File:Kyoto-Gel08301.jpg]]<br/>
-　We cultured σ54 promoter5<br/>
-'''Friday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Restriction of σ54 promoter5,glnL, glnG, glnL+G and rpoN<br/>
-　Cut them with EcoRl and Spel
-　After purification, the concentration of DNA were<br/>
-　σ54 promoter5: 23.6 ng/ul<br/>
-　glnL: 28.1 ng/ul<br/>
-　glnG: 26.3 ng/ul<br/>
-　glnL+G: 15.3 ng/ul<br/>
-　rpoN: 20.8 ng/ul<br/>
-・Ligation reaction<br/>
-　Ligated glnL, glnG and rpoN to pSB1K3.<br/>
-'''Thursday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Screening PCR of glnL, glnG, glnL+G and rpoN<br/>
-glnL
-　[[File:Kyoto-Gel09020.jpg]]<br/>
-glnG
-　[[File:Kyoto-Gel09021.jpg]]<br/>
-glnL+G & rpoN
-　[[File:Kyoto-Gel09022.jpg]]<br/>
-　We cultured glnL5, glnG4<br/>
-'''Saturday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Mini prep of glnL5 and glnG4<br/>
-　glnL5: 43.9 ng/ul<br/>
-　glnG4: 38.1 ng/ul<br/>
-・Screening PCR of rpoN
-　[[File:Kyoto-Gel09020.jpg]]<br/>
-Predation(Hashiya)<br/>
-・PCR amplification of glmS<br/>
-　--Primers--<br/>
-　left primer:ggaattcgcggccgcttctagagcaggttgaccgacaacgata<br/>
-　right primer:ggactagtacgcagggcatccatttat<br/>
-　[[File:Kyoto-Gel09030.jpg]]<br/>
-　lane1: 100bp DNA ladder, lane2: glmS from gDNA, lane3: glmS from ASKA clone<br/>
-・TA cloning of glmS<br/>
-　Ligated glmS from gDNA to pTA vector.<br/>
-'''Sunday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Screening PCR of rpoN<br/>
-　[[File:Kyoto-Gel09020.jpg]]<br/>
-・Transformation of bellow parts<br/>
--23L: BBa_B0015  (double terminator)<br/>
--18E: BBa_J23101 (constitutive promoter)<br/>
--18C: BBa_J23100 (constitutive promoter)<br/>
-<html><a name="lab-week6"></a></html>
-== Week6: Monday 5th September - Sunday 11th September ==
-'''Monday'''<br/>
-Nutritional Signal(Hashiya)<br/>
-・Screening PCR of glnL+G+double terminator<br/>
-'''Tuesday'''<br/>
-Luminescence(Kusaba, Hara):ハエの走行性実験②（②は改良版）　♂、緑×2回　♀、青×1回
-'''Wednesday'''<br/>
-Luminescence(Hara):ハエの走行性実験②　♂、青×2回　♀、緑×2回、青×1回
-'''Thursday'''<br/>
-Luminescence(Hara):ハエの走行性実験②　♂、紫外線×3回　♀、紫外線×3回
-'''Friday'''<br/>
-'''Saturday'''<br/>
-Luminescence(Kusaba, Hara):ハエの走行性実験②　♂、青×2回、赤×2回　♀、青×2回、赤×2回
-'''Sunday'''<br/>
-Luminescence(Kusaba, Hara):ハエの走行性実験②　♂、紫外線×2回、
-<html><a name="lab-week7"></a></html>
-== Week7: Monday 12th September - Sunday 18th September ==
-'''Monday'''<br/>
-Luminescence:大腸菌の形質転換(Hashiya)　ハエの走行性実験②(Kusaba, Hara)
-'''Tuesday'''<br/>
-Luminescence:大腸菌はじめて光る。しかし光量は少ない。
-'''Wednesday'''<br/>
-'''Thursday'''<br/>
-'''Friday'''<br/>
-Digestion(Kajita)
-PCR amplification of SAM-P20 and ChiA
-:We performed colony direct PCRs from a ''S. albogriseolus'' colony and a ''S. avermitilis'' colony.
-:{|class="wikitable"
-|+Reaction mixture
-! width="80"|Component
-!width="80"|Volume(&mu;l)
-|-
-|2x Buffer||25
-|-
-|2mM dNTPs||10
-|-
-|Primer 1||1.5
-|-
-|Primer 2||1.5
-|-
-|Template||X
-|-
-|ddH<sub>2</sub>O||up to 50
-|}
-:{|class="wikitable"
-|+PCR condition
-|Predenature||94C||2m||
-|-
-|Denature||98C||10s||rowspan="3"|25cycles
-|-
-|Annealing||56C||30s
-|-
-|Extension||68C||1m30s
-|}
-:We prepared two kinds of templates:
-#Picked a colony, suspended in 50ul of water and incubated 1min at 95 degree. Added 1ul to the reaction mixture.
-#Picked a colony and dipped in the reaction mixture.
-:Gel electrophoresis indicated that the PCR amplifications were successful for a sample, ChiA, but it was a faint band. We decided to retry direct PCR of SAM-P20 and PCR of ChiA.
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="lab-week8"></a></html>
-== Week8: Monday 19th September - Sunday 25th September ==
-'''Monday'''<br/>
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''Thursday'''<br/>
-'''Friday'''<br/>
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="lab-week9"></a></html>
-== Week9: Monday 26th September - Sunday 2nd October ==
-'''Monday'''<br/>
-'''Tuesday'''<br/>
-'''Wednesday'''<br/>
-'''Thursday'''<br/>
-'''Friday'''<br/>
-'''Saturday'''<br/>
-'''Sunday'''<br/>
-<html><a name="protocol"></a></html>
-== Protocol ==
-=== Medium for drosophila ===
-::{| class="wikitable"
-|+ [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila]
-! width="300" |Materials
-! width="300" |Methods
-|-
-|
-* water : 500mL
-* dry yeast : 20g
-* corn flour : 45g
-* glucose : 50g
-* agarose : 3.5~5g
-* propionic acid : 1.5mL
-* 10% p-hydroxybenzoate in 70% Eternol : 5g
-|
-# Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it.
-# Stir corn flour and glucose with the remaining water.
-# Stir 1 and 2, then autoclave it again.
-# after autoclave, add propionic acid and 10% p-hydroxybenzoate in 70% Eternol into it.■
-|}
 </div>
 <!-- /#main -->

Team:Kyoto/Notebook

From 2011.igem.org

Revision as of 12:01, 22 September 2011

Notebook