Difference between revisions of "Team:NYMU-Taipei/Notebook"

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Augest, 2017
+
August, 2017
 
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<p>
 
<p>
<b>2017-06-19</b>&nbsp;&nbsp; prepare stock1 and stock5<br>
+
<b>2017-08-04</b> &nbsp;&nbsp; Digest check: BBa_K112806
 +
<b>2017-08-05</b> &nbsp;&nbsp; <i>Chlorella Vulgaris</i> growth curve measuring
 +
<b>2017-08-06</b> &nbsp;&nbsp; PCR(Dream Taq): Sucide mechanism-BBa_K112806, K737007, J04450, K112000
 +
<b>2017-08-07</b> &nbsp;&nbsp; (1) PCR(KOD plus): Sucide mechanism-BBa_K112806, K737007, J04450, K112000
 +
<br>     (2) NrtA- annealing, PNK, ligation, transformation
 +
<b>2017-08-08</b> &nbsp;&nbsp; Suicide mechanism-Annealing, cloning, ligation, transformation
 +
<b>2017-08-09</b> &nbsp;&nbsp; Extraction of <i>Synechoccocus</i> PCC7942 genomic DNA
 +
<b>2017-08-10</b> &nbsp;&nbsp; (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
 +
<b>2017-08-11</b> &nbsp;&nbsp; (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
 +
<b>2017-08-12</b> &nbsp;&nbsp; (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
 +
<b>2017-08-13</b> &nbsp;&nbsp; (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
 +
<b>2017-08-14</b> &nbsp;&nbsp; (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
 +
<b>2017-08-15</b> &nbsp;&nbsp; (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
 +
<b>2017-08-16</b> &nbsp;&nbsp; (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
 +
<b>2017-08-18</b> &nbsp;&nbsp; (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation
 +
<br>     (2) PCR(KOD plus): PrbCL and indC
 +
<b>2017-08-19</b> &nbsp;&nbsp; (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation
 +
<br>     (2) Inoculate E. coli containing pBAD18
 +
<br>     (3) Fusion PCR: PrbCL+indC
 +
<b>2017-08-22</b> &nbsp;&nbsp; (1) PCR: Lycopene and MelA
 +
<br>     (2) Fusion PCR: PrbCL+indC
 +
<b>2017-08-24</b> &nbsp;&nbsp; (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
 +
<b>2017-08-25</b> &nbsp;&nbsp; (1) Endolysin- PCR, annealing, cloning, ligation, transformation
 +
<br>     (2) Liquid culture: PBR322
 +
<br>     (3) Liquid culture: B0015
 +
<b>2017-08-26</b> &nbsp;&nbsp; (1) Plasmid extraction: endolysin
 +
<br>     (2) Plasmid extraction: PBR322
 +
<br>     (3) Plasmid extraction: B0015
 +
<b>2017-08-27</b> &nbsp;&nbsp; (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
 +
<b>2017-08-28</b> &nbsp;&nbsp; (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
 +
<b>2017-08-29</b> &nbsp;&nbsp; (1) Nile red oil measuring of <i>Chlorella Vulgaris</i> (~0901)
 +
<br>     (2) Plasmid extraction: NrtA
 +
<br>     (3) Holin construction
 +
<br>     (4) sequencing: NrtA
 +
<b>2017-08-30</b> &nbsp;&nbsp; (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)
 +
<b>2017-08-31</b> &nbsp;&nbsp; (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%)
 
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September, 2017
 
September, 2017
 
</a>
 
</a>
 
<p>
 
<p>
<b>2017-06-19</b>&nbsp;&nbsp; prepare stock1 and stock5<br>
+
<b>2017-09-01</b> &nbsp;&nbsp; (1)Fusion PCR: Ampicillin resistance, B0015
 +
<br>     (2) Three pieces PCR: 5NSII, 3NSII, and PBR322 ori
 +
 
 +
<b>2017-09-02</b> &nbsp;&nbsp; (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
 +
 
 +
<b>2017-09-03</b> &nbsp;&nbsp; (1) Colony PCR: pPIGBACK
 +
<br>     (2) Liquid culture: pPIGBACK
 +
<br>     (3) PCR (KOD): CrtZ and PrbCL
 +
 
 +
<b>2017-09-04</b> &nbsp;&nbsp; (1) Plasmid extraction: pPIGBACK
 +
<br>     (2) Digestion check: pPIGBACK
 +
<br>     (3) Fusion PCR: CrtZ+ PrbCL
 +
 
 +
<b>2017-09-05</b> &nbsp;&nbsp; (1) pPIGBACK construct finish
 +
<br>     (2) Fusion PCR: PrbCL+IndC
 +
<br>     (3) Ligation: CrtZ+ PrbCL with pPIGBACK
 +
<br>     (4) Transformation: CrtZ construct
 +
 
 +
<b>2017-09-06</b> &nbsp;&nbsp; (1) Colony pcr: CrtZ construct
 +
<br>     (2) Liquid culture: CrtZ
 +
<br>     (3) Lycopene primers design
 +
<br>     (4) IndC primers redesign
 +
 
 +
<b>2017-09-07</b> &nbsp;&nbsp; (1) plasmid extraction: CrtZ-Ppigback
 +
<br>     (2) restriction enzyme check: CrtZ-pPIGBACK
 +
 
 +
<b>2017-09-08</b> &nbsp;&nbsp; (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%)
 +
 
 +
<b>2017-09-09</b> &nbsp;&nbsp; (1) nitrate and nitrite adsorption assay of Chlorella vulgaris (co-coculture with NrtA)
 +
<br>     (2) fusion PCR: PrbcL+indC
 +
 
 +
<b>2017-09-10</b> &nbsp;&nbsp; indC construct (~transformation)
 +
 
 +
<b>2017-09-19</b> &nbsp;&nbsp; (1) Endolysin-Holin construct
 +
<br>     (2) Site-directed primers design
 +
 
 +
<b>2017-09-20</b> &nbsp;&nbsp; (1) Liquid culture: BBa_J04450
 +
<br>     (2) Liquid culture: CrtZ-PCC7942 clones
 +
 
 +
<b>2017-09-21</b> &nbsp;&nbsp; Plasmid extraction: J04450, CrtZ-PCC7942
 +
 
 +
<b>2017-09-22</b> &nbsp;&nbsp; Site-directed PCR: 3NSII
 +
 
 +
<b>2017-09-23</b> &nbsp;&nbsp; Site-directed-3NSII construct finish
 +
 
 +
<b>2017-09-24</b> &nbsp;&nbsp; (1) plasmid extraction: Site-directed-3NSII
 +
<br>     (2) Site-directed PCR: 5NSII-1
 +
<br>     (3) Psb1c3-PrbcL construct finish
 +
 
 +
<b>2017-09-25</b> &nbsp;&nbsp; (1) Psb1c3-NrtA construct finish
 +
<br>     (2) colony PCR: Psb1c3-PrbcL
 +
<br>     (3) Liquid culture: Psb1c3-PrbcL
 +
<br>     (4) IndC construct
 +
 
 +
<b>2017-09-26</b> &nbsp;&nbsp; (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
 +
 
 +
<b>2017-09-27</b> &nbsp;&nbsp; (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
 +
 
 +
<b>2017-09-28</b> &nbsp;&nbsp; (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
 +
 
 +
<b>2017-09-29</b> &nbsp;&nbsp; (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
 +
 
 +
<b>2017-09-30</b> &nbsp;&nbsp; (1) Psb1c3-PrbcL(SD) finish
 +
<br>     (2) Liquid culture: CrtZ-pPIGBACK
 +
 
 
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Revision as of 08:44, 10 October 2017

Protocol

NrtA expression

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Suicide gene expression

Download the file

OD measurement

Download the file

Timeline