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<ul class="front"> | <ul class="front"> | ||
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design"><li class="list" id="project2">Design</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design"><li class="list" id="project2">Design</li></a> | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/Results"><li class="list" id="project3">Results</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Results"><li class="list" id="project3">Results</li></a> | ||
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate"><li class="list" id="project4">Demonstration</li></a> |
<a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab "><li class="list" id="project5">InterLab</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab "><li class="list" id="project5">InterLab</li></a> | ||
</ul> | </ul> | ||
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<li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a><hr> | <li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a><hr> | ||
<ul class="sub" id="sub_parts" style="cursor:default;"> | <ul class="sub" id="sub_parts" style="cursor:default;"> | ||
− | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part"><li class="list" id="parts1">Basic Part</li></a> | |
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part"><li class="list" id="parts2">Composite Part</li></a> |
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Improve"><li class="list" id="parts3">Improved Part</li></a> |
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Improve"><li class="list" id=" | + | |
</ul> | </ul> | ||
</li> | </li> | ||
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Model"><li class="list" id="model1">Overview</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Model"><li class="list" id="model1">Overview</li></a> | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/Binding"><li class="list" id="model2">Binding</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Binding"><li class="list" id="model2">Binding</li></a> | ||
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Polymer"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Polymer"><li class="list" id="model3">Polymer</li></a> |
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</ul> | </ul> | ||
</li> | </li> | ||
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Public_Engagement"><li class="list" id="human_practice2">Public Engagement</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Public_Engagement"><li class="list" id="human_practice2">Public Engagement</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/Entrepreneurship"><li class="list" id="human_practice3">Entrepreneurship</li></a> | ||
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/ | + | <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/Intergrate"><li class="list" id="human_practice5">Intergrated HP</li></a> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook"><li class="list" id="notebook1">Overview</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook"><li class="list" id="notebook1">Overview</li></a> | ||
<a href="https://2018.igem.org/Team:CCU_Taiwan/Collaborations"><li class="list" id="notebook2">Collaborations</li></a> | <a href="https://2018.igem.org/Team:CCU_Taiwan/Collaborations"><li class="list" id="notebook2">Collaborations</li></a> | ||
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Protocols"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Protocols"><li class="list" id="notebook3">Protocols</li></a> |
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments"><li class="list" id="notebook4">Experiments</li></a> |
− | <a href="https://2018.igem.org/Team:CCU_Taiwan/Materials"><li class="list" id=" | + | <a href="https://2018.igem.org/Team:CCU_Taiwan/Materials"><li class="list" id="notebook5">Materials</li></a> |
</ul> | </ul> | ||
</li> | </li> | ||
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</div> | </div> | ||
</nav> | </nav> | ||
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<div class="backgroundNotebook"> | <div class="backgroundNotebook"> | ||
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<div class="content"> | <div class="content"> | ||
Revision as of 12:07, 15 October 2018
Notebook
February
Molecular biology class
E-coli genomic DNA preparation
E-coli transformation
March
Instrument operation
April
Project brainstorming-Product Positioning, HGS monomer proportion
May
Culture selection-compare Yeast, E-coli, Acetobacter aceti
Gene design
June
Interlab experiment-Calbration1,2,3
July
7/3
Start interlab experiment-cell measurement
7/5
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
19:00-20:00
Day 2:
Pick 2 colonies from each of the transformation plates and inoculate in 5mL LB medium +Chloramphenicol. Grow the cells overnight (14 hours) at 37°C and 220 rpm.
8/6
10:00-18:00
Day 3: Cell growth, sampling, and assay
- Make a 1:10 dilution of each overnight culture in LB+Chloramphenicol (0.5mL of culture into 4.5mL of LB+Chlor)
- Measure Abs600 of these 1:10 diluted cultures
- Record the data in your notebook
- Dilute the cultures further to a target Abs600 of 0.02 in a final volume of 12 ml LB medium + Chloramphenicol in 50 mL falcon tube (amber, or covered with foil to block light).
- Take 500 μL samples of the diluted cultures at 0 hours into 1.5 ml eppendorf tubes, prior to incubation. (At each time point 0 hours and 6 hours, you will take a sample from each of the 8 devices, two colonies per device, for a total of 16 eppendorf tubes with 500 μL samples per time point, 32 samples total). Place the samples on ice.
- Measure your samples (Abs600 and Fluorescence measurement)
Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI,EcoRI
8/7
10:00-12:00
Fluorescence measurement
Ligation
8/8
Day 2:
Pick 2 colonies from each of the transformation plates and inoculate in 5mL LB medium +Chloramphenicol. Grow the cells overnight (16 hours) at 37°C and 220 rpm.
Transformation
- Add all pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (15µl) into 20µl ECOS™ 101 Competent Cells [DH5a]
- Incubate on ice 5 minutes
- Heat shock at 42℃ 45 second
- Incubate on ice 5 minutes
- Add 140µl LB
- Incubate at 37℃ 1hr
- Spread on LB+ Zeocin plate
Ligation
8/9
10:00-16:00
Day 3:Cell growth, sampling, and assay
- Make a 1:10 dilution of each overnight culture in LB+Chloramphenicol (0.5mL of culture into 4.5mL of LB+Chlor)
- Measure Abs600 of these 1:10 diluted cultures
- Record the data in your notebook
- Dilute the cultures further to a target Abs600 of 0.02 in a final volume of 12 ml LB medium + Chloramphenicol in 50 mL falcon tube (amber, or covered with foil to block light).
- Take 500 μL samples of the diluted cultures at 0 hours into 1.5 ml eppendorf tubes, prior to incubation. (At each time point 0 hours and 6 hours, you will take a sample from each of the 8 devices, two colonies per device, for a total of 16 eppendorf tubes with 500 μL samples per time point, 32 samples total). Place the samples on ice.
- Measure your samples (Abs600 and Fluorescence measurement)
- Spread on LB+ Zeocin plate
Transformation
- Add all pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (20µl) into 20µl Competent Cells DH5a and pGAPZ A 1µl+19µl ddH2O into 10µl Competent Cells DH5a as positive control.
- Incubate on ice 15min
- Heat shock at 42℃ 45sec
- Incubate on ice 5min
- Add 140µl LB
- Incubate at 37℃ 1hr
- Spread on LB+ Zeocin plate
8/10
10:00-16:00
Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI,EcoRI
Gel Extraction
8/11
Ligation
8/12
Transformation
pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
8/13
pGAPZ-A_Lac1-pGAPZ-A_Px16-pGAPZ-A_Px18-clone
20180813 PCR (pGAPZ-A_Lac1, pGAPZ-A_Px16, pGAPZ-A_Px18)
8/14
Miniprep
Result
8/15
12:00 Digest (use AgeI, EcoRI)
Protocol reference:
8/22
8/23
8/24
8/27
8/28
Transform
- pGAPZA-1 + His4 + Px16
- pGAPZA-1 + His4 + Px18
- pGAPZA-1 + His4 + Lac1
LB + zeocin
8/29
8/30
Check
pGAPZA-1 + His4
1. Digest - AgeI