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</p> | </p> | ||
<p class="second">8/5~8/12</p> | <p class="second">8/5~8/12</p> | ||
− | <p class="description">Cell growth, sampling, and assay<br> | + | <p class="description">    Cell growth, sampling, and assay<br> |
− | + |     Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<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™ 101 Competent Cells [DH5a]<br> |
− | + |     Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> | |
− | Transformation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (15µl) into 20µl ECOS™ 101 Competent Cells [DH5a]<br> | + |     Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<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> | ||
<p class="second">8/13~8/19</p> | <p class="second">8/13~8/19</p> | ||
− | <p class="description">Miniprep pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> | + | <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 | |
− | Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI | + | |
</p> | </p> | ||
<p class="second">8/20~8/26</p> | <p class="second">8/20~8/26</p> | ||
− | <p class="description">Gel extract<br> | + | <p class="description">  Gel extract<br> |
− | + |     Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | |
− | Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | + |     Ligation - pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 |
− | + | ||
− | Ligation - pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 | + | |
</p> | </p> | ||
<p class="second">8/27~9/2</p> | <p class="second">8/27~9/2</p> | ||
− | <p class="description">Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br><br> | + | <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> | |
− | Transform <br> | + |       2. pGAPZ A-1 + HIS4 + Px18<br> |
− | 1. pGAPZ A-1 + HIS4 + Px16 <br> | + |       3. pGAPZ A-1 + HIS4 + Lac1<br> |
− | 2. pGAPZ A-1 + HIS4 + Px18<br> | + |     Digest pGAPZ A HIS4(X2) with AgeI, EcoRI |
− | 3. pGAPZ A-1 + HIS4 + Lac1<br> | + | |
− | + | ||
− | + | ||
</p> | </p> | ||
<p class="second">9/3~9/9</p> | <p class="second">9/3~9/9</p> | ||
− | <p class="description">PCR—pGAPZ A-1 + HIS4 + Px16, Px18, Lac1<br> | + | <p class="description">    PCR—pGAPZ A-1 + HIS4 + Px16, Px18, Lac1<br> |
− | + |     miniprep – pGAPZ A-1 + HIS4 + Px16, Px18<br> | |
− | miniprep – pGAPZ A-1 + HIS4 + Px16, Px18<br> | + |     Transform – pGAPZ A-1 + HIS4 + Lac1<br> |
− | + |     Digestion – pGAPZ A-1 + HIS4+lac1<br> | |
− | Transform – pGAPZ A-1 + HIS4 + Lac1 | + | |
− | + | ||
− | + | ||
</p> | </p> | ||
<p class="second">9/12~9/14</p> | <p class="second">9/12~9/14</p> | ||
− | <p class="description">Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1<br> | + | <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> | |
− | + |     Plasmid miniprep - HIS4+Lac1, HIS4+Px16 | |
− | Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1<br> | + | |
− | + | ||
− | Plasmid miniprep - HIS4+Lac1, HIS4+Px16 | + | |
</p> | </p> | ||
<p class="second">9/24~9/27</p> | <p class="second">9/24~9/27</p> | ||
− | <p class="description">Culture HIS4+Lac1 x3<br> | + | <p class="description">    Culture HIS4+Lac1 x3<br> |
− | + |     Prepare yeast competent cells (SMD1168)<br> | |
− | + |     Traditional plasmid DNA extraction Day1 <br> | |
− | + |     Prepare YPD plates<br> | |
− | + |     Prepare the amino acids of SD plates<br> | |
− | Prepare YPD plates<br> | + |     Traditional plasmid DNA extraction Day2<br> |
− | Prepare the amino acids of SD plates<br> | + |     Plasmids contain HIS4 cut with SalI<br> |
− | + |     Yeast transformation - Px16, Px18, Lac1 (plasmids contain HIS4) | |
− | Traditional plasmid DNA extraction Day2<br> | + | |
− | Plasmids contain HIS4 cut with SalI<br> | + | |
− | + | ||
− | Yeast transformation - Px16, Px18, Lac1 (plasmids contain HIS4) | + | |
</p> | </p> | ||
<p class="second">10/8~10/11</p> | <p class="second">10/8~10/11</p> | ||
− | <p class="description">Prepare yeast competent cells (SMD1168)<br> | + | <p class="description">    Prepare yeast competent cells (SMD1168)<br> |
− | Colony PCR (HIS4+Px16)<br> | + |     Colony PCR (HIS4+Px16)<br> |
− | + |     Safety (30℃, 50℃, 70℃)<br> | |
− | Safety (30℃, 50℃, 70℃)<br> | + |     Yeast transformation - Px16, Lac1 (plasmids contain HIS4)<br> |
− | + |   Safety end | |
− | Yeast transformation - Px16, Lac1 (plasmids contain HIS4)<br> | + | |
− | + | ||
− | Safety end | + | |
</p> | </p> | ||
</div> | </div> |
Revision as of 11:01, 17 October 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)
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
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
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
Plasmid miniprep - HIS4+Lac1, HIS4+Px16
9/24~9/27
Culture HIS4+Lac1 x3
Prepare yeast competent cells (SMD1168)
Traditional plasmid DNA extraction Day1
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)
10/8~10/11
Prepare yeast competent cells (SMD1168)
Colony PCR (HIS4+Px16)
Safety (30℃, 50℃, 70℃)
Yeast transformation - Px16, Lac1 (plasmids contain HIS4)
Safety end