Difference between revisions of "Team:CCU Taiwan/Notebook"
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Medal"><li class="list" id="home3">Medals</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Medal"><li class="list" id="home3">Medals</li></a> | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/Judge"><li class="list" id="home4">For Judges</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Judge"><li class="list" id="home4">For Judges</li></a> | ||
| + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Achievements"><li class="list" id="home5">Achievements</li></a> | ||
</ul> | </ul> | ||
</li> | </li> | ||
| Line 130: | Line 131: | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship"><li class="list" id="human_practice3">Entrepreneurship</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship"><li class="list" id="human_practice3">Entrepreneurship</li></a> | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/engaging_experts"><li class="list" id="human_practice4">Engaging Experts</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/engaging_experts"><li class="list" id="human_practice4">Engaging Experts</li></a> | ||
| − | <a href="https://2018.igem.org/Team:CCU_Taiwan/ | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Integrate"><li class="list" id="human_practice5">Integrated HP</li></a> |
</ul> | </ul> | ||
</li> | </li> | ||
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</header> | </header> | ||
| + | <div class="indicator"> | ||
| + | |||
| + | <div class="pointerNotebook" id="1"><a href="#ca1">Notebook</a></div> | ||
| + | </div> | ||
<div class="backgroundNotebook"> | <div class="backgroundNotebook"> | ||
| − | <div class=" | + | <div class="photoNotebook"><h1 class="bigtitle">NOTEBOOK<h1></div> |
<div class="content"> | <div class="content"> | ||
| − | + | <p class="first" id="ca1">Notebook</p> | |
| + | <p class="second">February</p> | ||
| + | <p class="description">    Molecular biology class<br> | ||
| + |     <I>E. coli</I> genomic DNA preparation<br> | ||
| + |     <I>E. coli</I> transformation<br> | ||
| + | </p> | ||
| + | <p class="second">March</p> | ||
| + | <p class="description">    Instrument operation</p> | ||
| + | <p class="second">April</p> | ||
| + | <p class="description">    Project brainstorming: Product Positioning, Monolignol proportion</p> | ||
| + | <p class="second">May</p> | ||
| + | <p class="description">    Culture selection: compare Yeast, <I>E. coli</I>, Acetobacter aceti<br> | ||
| + |     Gene design | ||
| + | </p> | ||
| + | <p class="second">June</p> | ||
| + | <p class="description">    Interlab experiment: Calibration 1, 2, 3</p> | ||
| + | <p class="second">July</p> | ||
| + | <p class="description">  <strong>7/3</strong><br> | ||
| + |     Start Interlab experiment: cell measurement<br> | ||
| + |   <strong>7/5</strong><br> | ||
| + |     Yeast Extract–Peptone–Dextrose (YPD) formulation <br> | ||
| + |   <strong>7/9</strong><br> | ||
| + |     Yeast (X33) culture<br> | ||
| + |   <strong>7/13</strong><br> | ||
| + |     Fundraising briefing session<br> | ||
| + |   <strong>7/16</strong><br> | ||
| + |     Communicate with NCKU (Interlab & project)<br> | ||
| + |   <strong>7/18</strong><br> | ||
| + |     Communicate with BIT (project)<br> | ||
| + | </p> | ||
| + | <p class="second">August</p> | ||
| + | <p class="description"><strong>8/5~8/12</strong></p> | ||
| + | <p class="description">    Cell growth, sampling, and assay<br> | ||
| + |     Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<br> | ||
| + |     Transformation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (15µl) into 20µl ECOS™<br> | ||
| + |     101 Competent Cells [DH5a]<br> | ||
| + |     Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> | ||
| + |     Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<br> | ||
| + | </p> | ||
| + | <p class="description"><strong>8/13~8/19</strong></p> | ||
| + | <p class="description">    Miniprep pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> | ||
| + |     Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<br> | ||
| + |     Digest pGAPZ A with SalI<br> | ||
| + |     Incubate with DNA Polymerase I, Large (Klenow) Fragment remove SalI restriction site<br> | ||
| + |     Ligation pGAPZ A-SalI(pGAPZ A-1)<br> | ||
| + |     Transformation of pGAPZ A-SalI(pGAPZ A-1)<br> | ||
| + |     Minipreparation of pGAPZ A-SalI(pGAPZ A-1) | ||
| + | </p> | ||
| + | <p class="description"><strong>8/20~8/26</strong></p> | ||
| + | <p class="description">  Gel extract<br> | ||
| + |     Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | ||
| + |     Ligation - pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 | ||
| + | </p> | ||
| + | <p class="description"><strong>8/27~9/2</strong></p> | ||
| + | <p class="description">    Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | ||
| + |     Ligation – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br><br> | ||
| + | |||
| + |     Transform <br> | ||
| + |       1. pGAPZ A-1 + HIS4 + Px16 <br> | ||
| + |       2. pGAPZ A-1 + HIS4 + Px18<br> | ||
| + |       3. pGAPZ A-1 + HIS4 + Lac1<br> | ||
| + |     Digest pGAPZ A HIS4(X2) with AgeI, EcoRI<br> | ||
| + |     pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1 Traditional plasmid DNA extraction<br> | ||
| + |     Yeast SMD1168 Transformation(pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1) | ||
| + | </p> | ||
| + | |||
| + | <p class="second">September</p> | ||
| + | <p class="description"><strong>9/3~9/9</strong></p> | ||
| + | <p class="description">    PCR—pGAPZ A-1 + HIS4 + Px16, Px18, Lac1<br> | ||
| + |     miniprep – pGAPZ A-1 + HIS4 + Px16, Px18<br> | ||
| + |     Transform – pGAPZ A-1 + HIS4 + Lac1<br> | ||
| + |     Digestion – pGAPZ A-1 + HIS4+lac1<br> | ||
| + | </p> | ||
| + | <p class="description"><strong>9/12~9/14</strong></p> | ||
| + | <p class="description">    Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1<br> | ||
| + |     Culture HIS4+Px16, HIS4+Lac1<br> | ||
| + |     Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1<br> | ||
| + |     Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR <br> | ||
| + |     Plasmid miniprep - HIS4+Lac1, HIS4+Px16 | ||
| + | </p> | ||
| + | <p class="description"><strong>9/24~9/27</strong></p> | ||
| + | <p class="description">    Culture HIS4+Lac1 x3<br> | ||
| + |     Prepare yeast competent cells (SMD1168)<br> | ||
| + |     Traditional plasmid DNA extraction Day1 <br> | ||
| + |     Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR <br> | ||
| + |     Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot <br> | ||
| + |     Prepare YPD plates<br> | ||
| + |     Prepare the amino acids of SD plates<br> | ||
| + |     Traditional plasmid DNA extraction Day2<br> | ||
| + |     Plasmids contain HIS4 cut with SalI<br> | ||
| + |     Yeast transformation - Px16, Px18, Lac1 (plasmids contain HIS4) | ||
| + | </p> | ||
| + | |||
| + | <p class="second">October</p> | ||
| + | <p class="description"><strong>10/8~10/11</strong></p> | ||
| + | <p class="description">    Prepare yeast competent cells (SMD1168)<br> | ||
| + |     Colony PCR (HIS4+Px16)<br> | ||
| + |     Safety (30℃, 50℃, 70℃)<br> | ||
| + |     Yeast transformation - Px16, Lac1 (plasmids contain HIS4)<br><br> | ||
| + |     Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot <br> | ||
| + | <br><br> | ||
| + | </p> | ||
</div> | </div> | ||
</div> | </div> | ||
Latest revision as of 08:52, 1 December 2018
NOTEBOOK
Notebook
February
Molecular biology class
E. coli genomic DNA preparation
E. coli transformation
March
Instrument operation
April
Project brainstorming: Product Positioning, Monolignol proportion
May
Culture selection: compare Yeast, E. coli, Acetobacter aceti
Gene design
June
Interlab experiment: Calibration 1, 2, 3
July
7/3
Start Interlab experiment: cell measurement
7/5
Yeast Extract–Peptone–Dextrose (YPD) formulation
7/9
Yeast (X33) culture
7/13
Fundraising briefing session
7/16
Communicate with NCKU (Interlab & project)
7/18
Communicate with BIT (project)
August
8/5~8/12
Cell growth, sampling, and assay
Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
Transformation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (15µl) into 20µl ECOS™
101 Competent Cells [DH5a]
Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
8/13~8/19
Miniprep pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
Digest pGAPZ A with SalI
Incubate with DNA Polymerase I, Large (Klenow) Fragment remove SalI restriction site
Ligation pGAPZ A-SalI(pGAPZ A-1)
Transformation of pGAPZ A-SalI(pGAPZ A-1)
Minipreparation of pGAPZ A-SalI(pGAPZ A-1)
8/20~8/26
Gel extract
Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
Ligation - pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
8/27~9/2
Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
Ligation – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
Transform
1. pGAPZ A-1 + HIS4 + Px16
2. pGAPZ A-1 + HIS4 + Px18
3. pGAPZ A-1 + HIS4 + Lac1
Digest pGAPZ A HIS4(X2) with AgeI, EcoRI
pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1 Traditional plasmid DNA extraction
Yeast SMD1168 Transformation(pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1)
September
9/3~9/9
PCR—pGAPZ A-1 + HIS4 + Px16, Px18, Lac1
miniprep – pGAPZ A-1 + HIS4 + Px16, Px18
Transform – pGAPZ A-1 + HIS4 + Lac1
Digestion – pGAPZ A-1 + HIS4+lac1
9/12~9/14
Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1
Culture HIS4+Px16, HIS4+Lac1
Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR
Plasmid miniprep - HIS4+Lac1, HIS4+Px16
9/24~9/27
Culture HIS4+Lac1 x3
Prepare yeast competent cells (SMD1168)
Traditional plasmid DNA extraction Day1
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot
Prepare YPD plates
Prepare the amino acids of SD plates
Traditional plasmid DNA extraction Day2
Plasmids contain HIS4 cut with SalI
Yeast transformation - Px16, Px18, Lac1 (plasmids contain HIS4)
October
10/8~10/11
Prepare yeast competent cells (SMD1168)
Colony PCR (HIS4+Px16)
Safety (30℃, 50℃, 70℃)
Yeast transformation - Px16, Lac1 (plasmids contain HIS4)
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot