Difference between revisions of "Team:NYMU-Taipei/Notebook"
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{{NYMU-Taipei}} | {{NYMU-Taipei}} | ||
{{NYMU-Taipei-css}} | {{NYMU-Taipei-css}} | ||
| − | + | <html> | |
| − | <html | + | |
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<head> | <head> | ||
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<style> | <style> | ||
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body{ | body{ | ||
background-color:#f6ffb2; | background-color:#f6ffb2; | ||
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} | } | ||
/* Content of Notebook */ | /* Content of Notebook */ | ||
#Protocol{ | #Protocol{ | ||
| − | width: | + | width: 81%; |
| − | padding:2%; | + | padding: 2%; |
display:block; | display:block; | ||
float:right; | float:right; | ||
background-color:#f6ffb2; | background-color:#f6ffb2; | ||
font-family:'Fresca',sans-serif; | font-family:'Fresca',sans-serif; | ||
| − | + | font-size:20px; | |
| − | + | ||
} | } | ||
#Timeline{ | #Timeline{ | ||
| − | width: | + | width: 81%; |
| − | padding:2%; | + | padding: 2%; |
display:block; | display:block; | ||
float:right; | float:right; | ||
background-color:#cee1ea; | background-color:#cee1ea; | ||
font-family:'Fresca',sans-serif; | font-family:'Fresca',sans-serif; | ||
| − | + | font-size:18px; | |
| + | } | ||
#Labnote{ | #Labnote{ | ||
| − | width: | + | width: 81%; |
padding:2%; | padding:2%; | ||
display:block; | display:block; | ||
float:right; | float:right; | ||
| − | background-color:#ffeaea; | + | background-color: #ffeaea; |
font-family:'Fresca',sans-serif; | font-family:'Fresca',sans-serif; | ||
| + | font-size:20px; | ||
padding-bottom:250px; | padding-bottom:250px; | ||
| − | + | } | |
#Protocol h1 { | #Protocol h1 { | ||
| − | color: # | + | color: #205e1a; |
| − | font-size: | + | font-size: 40px; |
font-family: 'Acme', sans-serif; | font-family: 'Acme', sans-serif; | ||
} | } | ||
#Protocol h3 { | #Protocol h3 { | ||
| − | color: # | + | color: #3dba32; |
font-size: 28px; | font-size: 28px; | ||
| − | font-family: ' | + | font-family: 'Acme', sans-serif; |
| + | padding-left:20px; | ||
| + | } | ||
| + | |||
| + | #Protocol a { | ||
| + | color:#205e1a; | ||
| + | font-family: 'Acme', sans-serif; | ||
} | } | ||
#Timeline h1{ | #Timeline h1{ | ||
color: #1c3e4f; | color: #1c3e4f; | ||
| − | font-size: | + | font-size: 40px; |
font-family: 'Acme', sans-serif; | font-family: 'Acme', sans-serif; | ||
} | } | ||
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#Labnote h1{ | #Labnote h1{ | ||
color: #722525; | color: #722525; | ||
| − | font-size: | + | font-size: 40px; |
font-family: 'Acme', sans-serif; | font-family: 'Acme', sans-serif; | ||
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} | } | ||
| − | . | + | /* Show & hide */ |
| − | + | .expandable { | |
| − | + | /* overflow set to hidden to hide the expanded text */ | |
| − | + | overflow: hidden; | |
| − | + | /* all style changes will ease-in-out for 1s */ | |
| − | + | -moz-transition: all 1s ease-in-out; | |
| − | + | -ms-transition: all 1s ease-in-out; | |
| − | + | -o-transition: all 1s ease-in-out; | |
| + | -webkit-transition: all 1s ease-in-out; | ||
| + | transition: all 1s; | ||
} | } | ||
| − | . | + | .panel { |
| − | + | background-color: #fff; | |
| + | border: 1px solid transparent; | ||
| + | border-radius: 4px; | ||
| + | border-color: #ddd; | ||
| + | -webkit-box-shadow: 0 1px 1px rgba(0, 0, 0, .05); | ||
| + | box-shadow: 0 1px 1px rgba(0, 0, 0, .05); | ||
| + | padding:15px; | ||
| + | padding-top:20px; | ||
} | } | ||
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@media only screen and (max-width: 800px) { | @media only screen and (max-width: 800px) { | ||
.igem_2017_footer_wrapper {width:100%; margin-left:0px;} | .igem_2017_footer_wrapper {width:100%; margin-left:0px;} | ||
| − | #Protocol {width: | + | #Protocol {width:96%; margin-left:50px;} |
| − | #Timeline {width: | + | #Timeline {width:96%; margin-left:50px;} |
| − | #Labnote {width: | + | #Labnote {width:96%; margin-left:50px;} |
} | } | ||
| − | + | ||
</style> | </style> | ||
| − | + | <script> | |
| + | function toggleHeight1(e, maxHeight) { | ||
| + | e = document.getElementById("s1"); // e = the gray div | ||
| + | |||
| + | if(e.style.height != '24px') { | ||
| + | e.style.height = '24px'; // height of one line: 20px | ||
| + | } else { | ||
| + | e.style.height = maxHeight + 'px'; | ||
| + | } | ||
| + | } | ||
| + | |||
| + | function toggleHeight2(e, maxHeight) { | ||
| + | e = document.getElementById("s2"); // e = the gray div | ||
| + | |||
| + | if(e.style.height != '24px') { | ||
| + | e.style.height = '24px'; // height of one line: 20px | ||
| + | } else { | ||
| + | e.style.height = maxHeight + 'px'; | ||
| + | } | ||
| + | } | ||
| + | |||
| + | function toggleHeight3(e, maxHeight) { | ||
| + | e = document.getElementById("s3"); // e = the gray div | ||
| + | |||
| + | if(e.style.height != '24px') { | ||
| + | e.style.height = '24px'; // height of one line: 20px | ||
| + | } else { | ||
| + | e.style.height = maxHeight + 'px'; | ||
| + | } | ||
| + | } | ||
| + | function toggleHeight4(e, maxHeight) { | ||
| + | e = document.getElementById("s4"); // e = the gray div | ||
| + | |||
| + | if(e.style.height != '24px') { | ||
| + | e.style.height = '24px'; // height of one line: 20px | ||
| + | } else { | ||
| + | e.style.height = maxHeight + 'px'; | ||
| + | } | ||
| + | } | ||
| + | function toggleHeight5(e, maxHeight) { | ||
| + | e = document.getElementById("s5"); // e = the gray div | ||
| + | |||
| + | if(e.style.height != '24px') { | ||
| + | e.style.height = '24px'; // height of one line: 20px | ||
| + | } else { | ||
| + | e.style.height = maxHeight + 'px'; | ||
| + | } | ||
| + | } | ||
| + | </script> | ||
| + | |||
| + | </head> | ||
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<body> | <body> | ||
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| − | + | <div id="Timeline" > | |
| − | + | <center> | |
<h3></h3> | <h3></h3> | ||
<img src="https://static.igem.org/mediawiki/2017/f/f9/T--NYMU-Taipei--notebook.jpg" | <img src="https://static.igem.org/mediawiki/2017/f/f9/T--NYMU-Taipei--notebook.jpg" | ||
style="width:50%;border-radius:50%;border:5px solid #f6ffb2;"> | style="width:50%;border-radius:50%;border:5px solid #f6ffb2;"> | ||
| − | + | </center> | |
| − | + | <h3></h3> | |
| − | + | <h3></h3> | |
| − | + | <h1>Timeline</h1> | |
| − | + | <p></p> | |
| − | < | + | |
| − | + | <div class='panel'> | |
| + | <div id="s1" class="expandable" style='height: 24px;padding-top:5px;'> | ||
| + | <a href="#!" onclick="toggleHeight1(this, 270); return false" | ||
| + | style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;"> | ||
| + | June, 2017 | ||
| + | </a> | ||
| + | <p> | ||
| + | <br> | ||
| + | <b>2017-06-19</b> prepare stock1 and stock5<br> | ||
| + | <b>2017-06-20</b> (1) reprepare stock5<br> | ||
| + | (2) prepare BG11 medium<br> | ||
| + | (3) unfreeze <i>Synechocystis</i> sp. PCC6803 and start to cultivate in BG11<br> | ||
| + | <b>2017-06-29</b> (1) prepare Pseudomonas aeruginosa growing medium<br> | ||
| + | (2) activate Pseudomonas aeruginosa and start to cultivate in Petri dish<br> | ||
| + | (3) subculture <i>Synechocystis</i> sp. PCC6803 | ||
| + | </p> | ||
| + | |||
| + | <center> | ||
| + | <a href="#!" onclick="toggleHeight1(this, 270);" style='font-size:20px;color:#2c498c'> | ||
| + | click to close | ||
| + | </a> | ||
| + | </center> | ||
| + | </div> | ||
</div> | </div> | ||
| − | |||
| − | |||
| − | <div id=" | + | <div class='panel'> |
| − | < | + | <div id="s2" class="expandable" style='height: 24px;padding-top:5px;'> |
| − | + | <a href="#!" onclick="toggleHeight2(this, 1440); return false" | |
| − | + | style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;"> | |
| − | + | July, 2017 | |
| + | </a> | ||
| + | <p> | ||
| + | <br> | ||
| + | <b>2017-07-02</b> extract <i>Pseudomonas aeruginosa</i> plasmid and freeze into refrigerator<br> | ||
| + | <b>2017-07-03</b> proliferate <i>Pseudomonas aeruginosa</i> and freeze into refrigerator<br> | ||
| + | <b>2017-07-04</b> (1) prepare BG11 medium<br> | ||
| + | (2) proliferate <i>Chlorella vulgaris</i> in Petri dish and centrifuge tube<br> | ||
| + | <b>2017-07-05 to 07-10</b> cultivate <i>Synechocystis</i> sp. PCC6803 and <i>Chlorella vulgaris</i> in incubator<br> | ||
| + | <b>2017-07-11</b> (1) extract <i>Synechocystis</i> sp. PCC6803 plasmid<br> | ||
| + | (2) prepare LB containing ampicillin and chloramphenicol and prepare Petri dish<br> | ||
| + | <b>2017-07-12</b> transform BBa_K112806, BBa_K737007, BBa_K112000, BBa_J04450, BBa_K1356003, BBa_K1356004, and BBa_K356005 into competent cell<br> | ||
| + | <b>2017-07-13</b> (1) proliferate 07-12 transformed E.coli<br> | ||
| + | (2) proliferate <i>Chlorella vulgaris</i><br> | ||
| + | (3) proliferate <i>Synechocystis</i> sp. PCC6803<br> | ||
| + | (4) activate <i>Synechococcus elongatus</i> PCC7942<br> | ||
| + | <b>2017-07-14</b> (1)extract 07-13 proliferated E.coli plasmid<br> | ||
| + | (2) digestion: BBa_K112806, BBa_K737007, BBa_K112000, BBa_J04450, BBa_K1356003, BBa_K1356004, and BBa_K1356005<br> | ||
| + | (check the correction of transformed E.coli)<br> | ||
| + | <b>2017-07-17</b> (1) digestion again: BBa_K1356005, BBa_K112806, and BBa_K112000<br> | ||
| + | (2) retransform BBa_K112806 and BBa_K112000<br> | ||
| + | <b>2017-07-18</b> (1) proliferate BBa_K112806 transformed E.coli (BBa_K112000 failed)<br> | ||
| + | (2) subculture <i>Synechococcus elongatus</i> PCC7942<br> | ||
| + | (3) cultivate <i>Synechocystis</i> sp. PCC6803 in BG11<br> | ||
| + | <b>2017-07-19</b> (1) extract 07-18 BBa_K112806 transformed E.coli plasmid<br> | ||
| + | (2) digestion check<br> | ||
| + | (3) save BBa_K112806 transformed E.coli<br> | ||
| + | (4) save 07-18 <i>Synechocystis</i> sp. PCC6803<br> | ||
| + | <b>2017-07-20</b> (1) activate Rhizobium etli in PY medium<br> | ||
| + | (2) dilute primer 001~008<br> | ||
| + | (3) run NrtA and terminator PCR (failed)<br> | ||
| + | <b>2017-07-21 to 07-23</b> cultivate <i>Chlorella vulgaris</i> again for measuring growth curve<br> | ||
| + | <b>2017-07-24</b> (1) prepare <i>Rhizobium etli</i> liquid culture<br> | ||
| + | (2) activate <i>Synechococcus elongatus</i> PCC7942<br> | ||
| + | <b>2017-07-25</b> (1) extract <i>Synechocystis</i> sp. PCC6803 genomic DNA<br> | ||
| + | (2) run NrtA PCR<br> | ||
| + | <b>2017-07-26</b> (1) extract <i>Rhizobium etli</i> genomic DNA<br> | ||
| + | (2) run NrtA and terminator PCR<br> | ||
| + | (3) run NrtA+ terminator fusion PCR (failed)<br> | ||
| + | <b>2017-07-27</b> (1) run NrtA+ terminator fusion PCR again<br> | ||
| + | (2) purify the successful fusion product <br> | ||
| + | (3) digestion check<br> | ||
| + | <b>2017-07-28</b> run the whole NrtA cloning procedure again, using KOD polymerase<br> | ||
| + | (1) run NrtA and terminator PCR<br> | ||
| + | (2) digestion check NrtA and terminator and purify the PCR product<br> | ||
| + | (3) run NrtA and terminator fusion PCR<br> | ||
| + | (4) digestion check and purify the fusion PCR product (failed)<br> | ||
| + | <b>2017-07-29</b> (1) run NrtA and terminator fusion PCR again<br> | ||
| + | (2) digestion check and purify the fusion PCR product<br> | ||
| + | (3) ligation: backbone(psb1C3) and promoter, RBS, NrtA, and terminator<br> | ||
| + | (4) transformation in to E.coli<br> | ||
| + | <b>2017-07-30</b> cultivate the transformed E.coli in liquid culture<br> | ||
| + | <b>2017-07-31</b> (1) extract 07-30 transformed E.coli plasmid<br> | ||
| + | (2) restriction enzyme check (failed)<br> | ||
| + | </p> | ||
| − | + | <center> | |
| − | + | <a href="#!" onclick="toggleHeight2(this,1440);" style='font-size:20px;color:#2c498c'> | |
| − | + | click to close | |
| − | + | </a> | |
| − | + | </center> | |
| − | + | </div> | |
| − | + | </div> | |
| − | + | ||
| − | + | <div class='panel'> | |
| − | + | <div id="s3" class="expandable" style='height: 24px;padding-top:5px;'> | |
| − | + | <a href="#!" onclick="toggleHeight3(this, 2200); return false" | |
| − | + | style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;"> | |
| − | + | August, 2017 | |
| − | + | </a> | |
| − | + | <p> | |
| − | + | <br> | |
| − | + | <b>2017-08-04</b> Digest check: BBa_K112806<br> | |
| − | + | <b>2017-08-05</b> <i>Chlorella Vulgaris</i> growth curve measuring<br> | |
| − | + | <b>2017-08-06</b> PCR(Dream Taq): Sucide mechanism-BBa_K112806, K737007, J04450, K112000<br> | |
| − | < | + | <b>2017-08-07</b> (1) PCR(KOD plus): Sucide mechanism-BBa_K112806, K737007, J04450, K112000 |
| − | + | <br> (2) NrtA- annealing, PNK, ligation, transformation<br> | |
| − | + | <b>2017-08-08</b> Suicide mechanism-Annealing, cloning, ligation, transformation<br> | |
| − | + | <b>2017-08-09</b> Extraction of <i>Synechoccocus</i> PCC7942 genomic DNA<br> | |
| − | + | <b>2017-08-10</b> (1) <i>Chlorella Vulgaris</i> growth curve measuring | |
| − | + | <br> (2) Fusion PCR: NrtA + terminator | |
| − | + | <br> (3) Digestion check: NrtA cut with BamH1 | |
| − | + | <br> (4) Subculture <i>S.elongatus</i> PCC7942<br> | |
| − | + | <b>2017-08-11</b> (1) Ligation: NrtA + terminator + promoter + RBS | |
| − | + | <br> (2) Ligation: BBa_K737007+K112806, J04450+K112000 (Endolysin and Holin) | |
| − | + | <br> (3) Transformation of Endolysin and Holin | |
| − | + | <br> (4) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i> | |
| − | + | <br> (5) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br> | |
| − | + | <b>2017-08-12</b> (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i> | |
| − | + | <br> (2) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br> | |
| − | + | <b>2017-08-13</b> (1) Transformation: BBa_K274002 and K1152008 into E. coli | |
| − | + | <br> (2) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i> | |
| − | + | <br> (3) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br> | |
| − | + | <b>2017-08-14</b> (1) Colony PCR: K1152008 | |
| − | + | <br> (2) Suicide mechanism- PCR, annealing, cloning, ligation, transformation | |
| − | + | <br> (3) NrtA-PCR, annealing, cloning, ligation, transformation | |
| − | + | <br> (4) Liquid culture-K1152008<br> | |
| − | + | <b>2017-08-15</b> (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i> | |
| − | + | <br> (2) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942 | |
| − | + | <br> (3) Suicide mechanism- final annealing, cloning, ligation, transformation | |
| − | + | <br> (4) Plasmid extraction and RE check: k1152008<br> | |
| − | + | <b>2017-08-16</b> (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i> | |
| − | + | <br> (2) OD value measurement of Chlorella Vulgaris and <i>S.elongatus</i> PCC7942 | |
| − | + | <br> (3) PCR(Dream Taq): PrbCL and indC<br> | |
| − | + | <b>2017-08-18</b> (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation | |
| − | + | <br> (2) PCR(KOD plus): PrbCL and indC<br> | |
| − | + | <b>2017-08-19</b> (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation | |
| − | + | <br> (2) Inoculate E. coli containing pBAD18 | |
| − | + | <br> (3) Fusion PCR: PrbCL+indC<br> | |
| − | + | <b>2017-08-22</b> (1) PCR: Lycopene and MelA | |
| − | + | <br> (2) Fusion PCR: PrbCL+indC<br> | |
| − | + | <b>2017-08-24</b> (1) Liquid culture: BBa_K274002 | |
| − | + | <br> (2) Extract PCC7942 genomic DNA | |
| − | + | <br> (3) Inoculate E. coli containing PBR322 | |
| − | + | <br> (4) Transformation: BBa_B0015 into E. coli<br> | |
| − | + | <b>2017-08-25</b> (1) Endolysin- PCR, annealing, cloning, ligation, transformation | |
| − | + | <br> (2) Liquid culture: PBR322 | |
| − | + | <br> (3) Liquid culture: B0015<br> | |
| − | + | <b>2017-08-26</b> (1) Plasmid extraction: endolysin | |
| − | + | <br> (2) Plasmid extraction: PBR322 | |
| − | + | <br> (3) Plasmid extraction: B0015<br> | |
| − | + | <b>2017-08-27</b> (1) PCR: Lycopene and MelA | |
| − | + | <br> (2) Digestion check: PBR322 | |
| − | + | <br> (3) Digestion check: B0015 | |
| − | + | <br> (4) pigment backbone primer design | |
| − | + | <br> (5) PCR (Dream Taq): CrtZ | |
| − | + | <br> (6) Plasmid extraction: endolysin | |
| − | + | <br> (7) Digestion check: endolysin<br> | |
| − | + | <b>2017-08-28</b> (1) Colony PCR: NrtA | |
| − | + | <br> (2) Liquid culture: NrtA | |
| − | + | <br> (3) Sequencing: endolysin | |
| − | + | <br> (4) Fusion PCR: PrbcL+IndC | |
| − | + | <br> (5) PCR(Dream taq): Holin(kit) | |
| − | + | <br> (6) Check pBR322 concentration and transform pBR322<br> | |
| − | + | <b>2017-08-29</b> (1) Nile red oil measuring of <i>Chlorella Vulgaris</i> (~0901) | |
| − | + | <br> (2) Plasmid extraction: NrtA | |
| − | + | <br> (3) Holin construction | |
| − | + | <br> (4) sequencing: NrtA<br> | |
| − | + | <b>2017-08-30</b> (1) PCR (Dream Taq): Ampicillin resistance, 5NSII, 3NSII, B0015, and PBR322 ori | |
| − | < | + | <br> (2) Endolysin construct finish |
| − | + | <br> (3) Plasmid extraction: pBR322 | |
| − | < | + | <br> (4) Restriction enzyme check: pBR322 |
| − | + | <br> (5) Backbone construct-3NSII, 5NSII,AMP, B0015, and ORI (PCR: dream taq)<br> | |
| − | + | <b>2017-08-31</b> (1) PCR (KOD plus): Ampicillin, 5NSII, 3NSII, B0015, and PBR322 ori | |
| − | + | <br> (2) PCR (Dream Taq): PBR322 ori and B0015 | |
| − | + | <br> (3) pSB1C3-NrtA construct finish | |
| − | + | <br> (4) Backbone construct-3NSII, 5NSII, AMP, B0015, and ORI (PCR: KOD) | |
| − | + | <br> (5) Transform: BBa_K112000 | |
| − | + | <br> (6) LBA: glucose + CM (0.3%) | |
| − | + | </p> | |
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| − | </ | + | |
| + | <center> | ||
| + | <a href="#!" onclick="toggleHeight3(this, 2200);" style='font-size:20px;color:#2c498c'> | ||
| + | click to close | ||
| + | </a> | ||
| + | </center> | ||
| + | </div> | ||
</div> | </div> | ||
| − | <div id=" | + | <div class='panel'> |
| − | < | + | <div id="s4" class="expandable" style='height: 24px;padding-top:5px;'> |
| − | < | + | <a href="#!" onclick="toggleHeight4(this, 1900); return false" |
| − | + | style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;"> | |
| − | + | September, 2017 | |
| + | </a> | ||
| + | <p> | ||
| + | <br> | ||
| + | <b>2017-09-01</b> (1)Fusion PCR: Ampicillin resistance, B0015 | ||
| + | <br> (2) Three pieces PCR: 5NSII, 3NSII, and PBR322 ori<br> | ||
| + | |||
| + | <b>2017-09-02</b> (1) Ligation: Ampicillin resistance, B0015+ 5NSII, 3NSII, and PBR322 ori | ||
| + | <br> (2) Transformation: pPIGBACK (pigment plasmid backbone) | ||
| + | <br> (3) PCR (Dream Taq): CrtZ and PrbCL<br> | ||
| + | |||
| + | <b>2017-09-03</b> (1) Colony PCR: pPIGBACK | ||
| + | <br> (2) Liquid culture: pPIGBACK | ||
| + | <br> (3) PCR (KOD): CrtZ and PrbCL<br> | ||
| + | |||
| + | <b>2017-09-04</b> (1) Plasmid extraction: pPIGBACK | ||
| + | <br> (2) Digestion check: pPIGBACK | ||
| + | <br> (3) Fusion PCR: CrtZ+ PrbCL<br> | ||
| + | |||
| + | <b>2017-09-05</b> (1) pPIGBACK construct finish | ||
| + | <br> (2) Fusion PCR: PrbCL+IndC | ||
| + | <br> (3) Ligation: CrtZ+ PrbCL with pPIGBACK | ||
| + | <br> (4) Transformation: CrtZ construct<br> | ||
| + | |||
| + | <b>2017-09-06</b> (1) Colony pcr: CrtZ construct | ||
| + | <br> (2) Liquid culture: CrtZ | ||
| + | <br> (3) Lycopene primers design | ||
| + | <br> (4) IndC primers redesign<br> | ||
| + | |||
| + | <b>2017-09-07</b> (1) plasmid extraction: CrtZ-Ppigback | ||
| + | <br> (2) restriction enzyme check: CrtZ-pPIGBACK<br> | ||
| + | |||
| + | <b>2017-09-08</b> (1) transform CrtZ-Ppigback into PCC7942 | ||
| + | <br> (2) nitrate and nitrite adsorption assay of Chlorella vulgaris (co-coculture with NrtA) | ||
| + | <br> (3) transform suicide mechanism construct (LBA+glucose 0.3%)<br> | ||
| + | |||
| + | <b>2017-09-09</b> (1) nitrate and nitrite adsorption assay of Chlorella vulgaris (co-coculture with NrtA) | ||
| + | <br> (2) fusion PCR: PrbcL+indC<br> | ||
| + | |||
| + | <b>2017-09-10</b> indC construct (~transformation)<br> | ||
| + | |||
| + | <b>2017-09-19</b> (1) Endolysin-Holin construct | ||
| + | <br> (2) Site-directed primers design<br> | ||
| + | |||
| + | <b>2017-09-20</b> (1) Liquid culture: BBa_J04450 | ||
| + | <br> (2) Liquid culture: CrtZ-PCC7942 clones<br> | ||
| + | |||
| + | <b>2017-09-21</b> Plasmid extraction: J04450, CrtZ-PCC7942<br> | ||
| + | |||
| + | <b>2017-09-22</b> Site-directed PCR: 3NSII<br> | ||
| + | |||
| + | <b>2017-09-23</b> Site-directed-3NSII construct finish<br> | ||
| + | |||
| + | <b>2017-09-24</b> (1) plasmid extraction: Site-directed-3NSII | ||
| + | <br> (2) Site-directed PCR: 5NSII-1 | ||
| + | <br> (3) Psb1c3-PrbcL construct finish<br> | ||
| + | |||
| + | <b>2017-09-25</b> (1) Psb1c3-NrtA construct finish | ||
| + | <br> (2) colony PCR: Psb1c3-PrbcL | ||
| + | <br> (3) Liquid culture: Psb1c3-PrbcL | ||
| + | <br> (4) IndC construct<br> | ||
| + | |||
| + | <b>2017-09-26</b> (1) Plasmid extraction and restriction enzyme check: Site-directed-5NSII-1 | ||
| + | <br> (2) Liquid culture: Psb1c3-NrtA | ||
| + | <br> (3) Colony PCR: IndC | ||
| + | <br> (4) Psb1c3-3NSII construct finish | ||
| + | <br> (5) Site-directed PCR: 5NSII-2<br> | ||
| + | |||
| + | <b>2017-09-27</b> (1) Psb1c3-NrtA construct finish | ||
| + | <br> (2) Plasmid extraction and restriction enzyme check: IndC | ||
| + | <br> (3) Plasmid extraction and restriction enzyme check: IndC | ||
| + | <br> (4) Psb1c3-3NSII construct finish Plasmid extraction and RE check: Site-directed 5NSII-2 | ||
| + | <br> (5) Site-directed PCR: AmpR<br> | ||
| + | |||
| + | <b>2017-09-28</b> (1) Plasmid extraction and RE check: Site-directed-AmpR | ||
| + | <br> (2) Plasmid extraction and RE check: Psb1c3-5NSII | ||
| + | <br> (3) Psb1c3-3NSII(SD) finish | ||
| + | <br> (4) Liquid culture: Psb1c3-PrbcL<br> | ||
| + | |||
| + | <b>2017-09-29</b> (1) Psb1c3-5NSII construct finish | ||
| + | <br> (2) Psb1c3-5NSII-ORI-3NSII construct finish | ||
| + | <br> (3) Colony PCR: Psb1c3-PrbcL- IndC | ||
| + | <br> (4) Colony PCR: Psb1c3-PrbcL- IndC | ||
| + | <br> (5) Psb1c3-PrbcL(SD) construct finish | ||
| + | <br> (6) Psb1c3-PrbcL-IndC construct finish | ||
| + | <br> (7) Site-directed: amp finish | ||
| + | <br> (8) transform CrtZ-pPIGBACK into PCC7942<br> | ||
| + | |||
| + | <b>2017-09-30</b> (1) Psb1c3-PrbcL(SD) finish | ||
| + | <br> (2) Liquid culture: CrtZ-pPIGBACK | ||
| + | |||
| + | </p> | ||
| + | |||
| + | <center> | ||
| + | <a href="#!" onclick="toggleHeight4(this, 1900);" style='font-size:20px;color:#2c498c'> | ||
| + | click to close | ||
| + | </a> | ||
| + | </center> | ||
</div> | </div> | ||
| − | + | </div> | |
| − | + | ||
| + | <div class='panel'> | ||
| + | <div id="s5" class="expandable" style='height: 24px;padding-top:5px;'> | ||
| + | <a href="#!" onclick="toggleHeight5(this, 2240); return false" | ||
| + | style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;"> | ||
| + | October, 2017 | ||
| + | </a> | ||
| + | <p> | ||
| + | <br> | ||
| + | <b>2017-10-01</b> (1) Colony PCR: 5NSII-Psb1c3 | ||
| + | <br> (2) Liquid culture: 5NSII-Psb1c3 | ||
| + | <br> (3) Colony PCR: 5NSII-ORI-3NSII-Psb1c3 | ||
| + | <br> (4) Liquid culture: Site-directed mutated clones | ||
| + | <br> (5) Construct: Psb1c3-PrbcL-IndC | ||
| + | <br> (6) Liquid culture: NrtA-C33 | ||
| + | <br><b>2017-10-02</b> (1) Finish all aite-directed mutagenesis | ||
| + | <br> (2) plasmid extraction: Psb1c3-5NSII, 5NSII-ORI-3NSII-Psb1c3 | ||
| + | <br> (3) Digestion check: Psb1c3-5NSII, 5NSII-ORI-3NSII-Psb1c3 | ||
| + | <br> (4) Colony PCR: Psb1c3-PrbcL-IndC | ||
| + | <br> (5) Liquid culture: Psb1c3-PrbcL-IndC | ||
| + | <br> (6) Liquid culture (French Press): NrtA-C33, competent cell | ||
| + | |||
| + | <br><b>2017-10-03</b> (1) French Press and dialysis | ||
| + | <br> (2) Sequencing: MelA | ||
| + | <br> (3) Construct Psb1c3-NrtA-Endolysin | ||
| + | |||
| + | <br><b>2017-10-04</b> (1) Construct pPIGBACK-J04450 | ||
| + | <br> (2) Construct Psb1c3-B0015-AMP | ||
| + | <br> (3) Construct Psb1c3-PrbcL-CrtZ | ||
| + | <br> (4) check nitrate concentration | ||
| + | <br> (5) Colony PCR: NrtA-endolysin | ||
| + | <br> (6) Liquid culture: NrtA-endolysin | ||
| + | |||
| + | <br><b>2017-10-05</b> (1) Colony PCR: pPIGBACK-J04450 | ||
| + | <br> (2) Liquid culture: pPIGBACK-J04450 | ||
| + | <br> (3) Digestion check: NrtA-endolysin | ||
| + | |||
| + | <br><b>2017-10-06</b> (1) DNA extraction and digestion-pPIGBACK-CrtZ | ||
| + | <br> (2) Finish construct pPIGBACK-J04450 | ||
| + | <br> (3) Colony PCR: Psb1c3-B0015-AMP | ||
| + | <br> (4) Liquid culture: Psb1c3-B0015-AMP | ||
| + | |||
| + | <br><b>2017-10-07</b> (1) Plasmid extraction: Psb1c3-B0015-AMP | ||
| + | <br> (2) Digest check: Psb1c3-B0015-AMP | ||
| + | <br> (3) Construct: Psb1c3-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-08</b> (1) Colony PCR: Psb1c3-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-09</b> (1) Liquid culture: competent cell for electroporation | ||
| + | <br> (2) Frech Press | ||
| + | <br> (3) Nitrate concentration assay | ||
| + | |||
| + | <br><b>2017-10-10</b> (1) Electroporation: competent cell | ||
| + | |||
| + | <br><b>2017-10-11</b> (1) Test electroporation: Psb1c3-J04450 | ||
| + | |||
| + | <br><b>2017-10-13</b> (1) Construct: Psb1c3-IndC (electroporation) | ||
| + | <br> (2) Construct: pPIGBACK-IndC (electroporation) | ||
| + | <br> (3) Construct: pPIGBACK-PrbcL-CrtZ | ||
| + | <br> (4) Construct: Psb1c3-B0015-AMP | ||
| + | <br> (5) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | <br> (6) Suicide mechanism functional test | ||
| + | |||
| + | <br><b>2017-10-14</b> (1) Colony PCR: pPIGBACK-IndC | ||
| + | <br> (2) Liquid culture: pPIGBACK-IndC | ||
| + | <br> (3) Colony PCR: pPIGBACK-PrbcL-CrtZ | ||
| + | <br> (4) <i>Chlorella Vulgaris</i> oil functional test: NrtA | ||
| + | <br> (5) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | <br> (6) Suicide mechanism functional test | ||
| + | |||
| + | <br><b>2017-10-15</b> (1) Finish construct pPIGBACK-PrbcL-IndC, Psb1c3-PrbcL-IndC | ||
| + | <br> (2) Liquid culture: All parts | ||
| + | <br> (3) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-16</b> (1) <i>Chlorella Vulgaris</i> oil functional test: NrtA | ||
| + | <br> (2) Plasmid extraction: all parts | ||
| + | <br> (3) Digestion check: all parts | ||
| + | <br> (4) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-17</b> (1) NrtA add to device again | ||
| + | <br> (2) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-18</b> (1) NrtA add to device again | ||
| + | <br> (2) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-19</b> (1) NrtA add to device again | ||
| + | <br> (2) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-20</b> (1) NrtA add to device again | ||
| + | <br> (2) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-21</b> (1) NrtA add to device again | ||
| + | <br> (2) Starch functional test: pPIGBACK-PrbcL-CrtZ | ||
| + | |||
| + | <br><b>2017-10-22</b> (1) PCR: MelA, IndC | ||
| + | <br> (2) Ligation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC | ||
| + | <br> (3) Transformation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC | ||
| + | |||
| + | <br><b>2017-10-23</b> (1) PCR: MelA, IndC, Psb1c3-B0015-AMP | ||
| + | <br> (2) Ligation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC, Psb1c3-B0015-AMP | ||
| + | <br> (3) Liquid culture: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC | ||
| + | |||
| + | <br><b>2017-10-24</b> (1) PCR: PrbcL-MelA, Psb1c3-IndC, pPIGBACK-IndC | ||
| + | <br> (2) Ligation: Psb1c3-PrbcL-MelA, pPIGBACK-PrbcL-MelA | ||
| + | <br> (3) Transformation: Psb1c3-PrbcL-MelA, pPIGBACK-PrbcL-MelA, Psb1c3-IndC, pPIGBACK-IndC, Psb1c3-B0015-AMP | ||
| + | <br> (4) Liquid culture: Psb1c3-B0015-AMP | ||
| + | |||
| + | <br><b>2017-10-25</b> (1) Digestion check all parts | ||
| + | <br> (2) Plasmid extraction: Psb1c3-B0015-AMP | ||
| + | <br> (3) Digestion check: Psb1c3-B0015-AMP | ||
| + | |||
| + | </p> | ||
| + | |||
| + | <center> | ||
| + | <a href="#!" onclick="toggleHeight5(this, 2240);" style='font-size:20px;color:#2c498c'> | ||
| + | click to close | ||
| + | </a> | ||
| + | </center> | ||
| + | </div> | ||
| + | </div> | ||
| + | |||
| + | </div> | ||
| + | |||
| + | <div id="Labnote"> | ||
| + | <h1>Labnote</h1> | ||
| + | <p></p> | ||
| + | <a href="https://static.igem.org/mediawiki/2017/0/04/T--NYMU-Taipei--labnote.pdf" target="_blank">Download the file</a> | ||
| + | <p></p> | ||
| + | <embed src="https://static.igem.org/mediawiki/2017/0/04/T--NYMU-Taipei--labnote.pdf" style="width:80%;height:500px;padding-left:10%"> | ||
| + | </div> | ||
</body> | </body> | ||
</html> | </html> | ||
Latest revision as of 16:14, 28 November 2017
