Difference between revisions of "Team:Tec-Monterrey/Notebook"

 
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                                 <li><a href="https://2016.igem.org/Team:Tec-Monterrey/Description">Project</a></li>
 
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                             <p>Human Practice</p>
 
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                                 <li><a href="https://2016.igem.org/Team:Tec-Monterrey/Notebook">Notebook</a></li>
 
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                     <h1 class="sep27" style="display: none;">Tuesday, September 27, 2016</h1>
 
                     <h1 class="sep27" style="display: none;">Tuesday, September 27, 2016</h1>
 
                     <h1 class="sep30" style="display: none;">Friday, September 30, 2016</h1>
 
                     <h1 class="sep30" style="display: none;">Friday, September 30, 2016</h1>
                     <h1 class="oct01" style="display: none;">Saturday, October 01, 2016</h1>
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                     <h1 class="oct1" style="display: none;">Saturday, October 01, 2016</h1>
                     <h1 class="oct02" style="display: none;">Sunday, October 02, 2016</h1>
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                     <h1 class="oct2" style="display: none;">Sunday, October 02, 2016</h1>
                     <h1 class="oct03" style="display: none;">Monday, October 03, 2016</h1>
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                     <h1 class="oct3" style="display: none;">Monday, October 03, 2016</h1>
                     <h1 class="oct04" style="display: none;">Tuesday, October 04, 2016</h1>
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                     <h1 class="oct4" style="display: none;">Tuesday, October 04, 2016</h1>
                     <h1 class="oct05" style="display: none;">Wednesday, October 05, 2016</h1>
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                     <h1 class="oct5" style="display: none;">Wednesday, October 05, 2016</h1>
                     <h1 class="oct06" style="display: none;">Thursday, October 06, 2016</h1>
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                     <h1 class="oct6" style="display: none;">Thursday, October 06, 2016</h1>
                     <h1 class="oct07" style="display: none;">Friday, October 07, 2016</h1>
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                     <h1 class="oct7" style="display: none;">Friday, October 07, 2016</h1>
                     <h1 class="oct08" style="display: none;">Saturday, October 08, 2016</h1>
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                     <h1 class="oct8" style="display: none;">Saturday, October 08, 2016</h1>
                     <h1 class="oct09" style="display: none;">Sunday, October 09, 2016</h1>
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                     <h1 class="oct9" style="display: none;">Sunday, October 09, 2016</h1>
 
                     <h1 class="oct10" style="display: none;">Monday, October 10, 2016</h1>
 
                     <h1 class="oct10" style="display: none;">Monday, October 10, 2016</h1>
 
                     <h1 class="oct11" style="display: none;">Tuesday, October 11, 2016</h1>
 
                     <h1 class="oct11" style="display: none;">Tuesday, October 11, 2016</h1>
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                     <h3 class="jun30">We tested our antibiotics: ampicillin, kanamycin, chloramphenicol with our PLYS and Shuffle bacterias</h3>
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                     <h3 class="jun30">We tested our antibiotics: ampicillin, kanamycin, chloramphenicol with our PLYS and Shuffle bacteria</h3>
 
                     <h3 class="jul2" style="display: none;">We repeated the testing of our antibiotics and we add BL21 to the experiment.</h3>  
 
                     <h3 class="jul2" style="display: none;">We repeated the testing of our antibiotics and we add BL21 to the experiment.</h3>  
 
                     <h3 class="jul4" style="display: none;">Transformation (Shuffle calcium competent E.coli). The parts used were:  GFP [AMP] (11P from the 2014 IGEM kit plate #4), Promotor (17P from the 2015 IGEM kit plate #4). There was no growth at all (even in our control). Probably the heat shock wasn’t done correctly (1.20 seconds, 42°c instead 0.50 s) . We reactivated our strains Top10 and DH5-alpha.</h3>  
 
                     <h3 class="jul4" style="display: none;">Transformation (Shuffle calcium competent E.coli). The parts used were:  GFP [AMP] (11P from the 2014 IGEM kit plate #4), Promotor (17P from the 2015 IGEM kit plate #4). There was no growth at all (even in our control). Probably the heat shock wasn’t done correctly (1.20 seconds, 42°c instead 0.50 s) . We reactivated our strains Top10 and DH5-alpha.</h3>  
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                     <h3 class="sep27" style="display: none;">We took out from the iGEM plates the following parts: 20F from 2015 plate and 13K from 2012 plate.</h3>
 
                     <h3 class="sep27" style="display: none;">We took out from the iGEM plates the following parts: 20F from 2015 plate and 13K from 2012 plate.</h3>
 
                     <h3 class="sep30" style="display: none;">An inoculum of C. violaceum was left to make electrocompetent cells. Also, a test of antibiotic resistance was made in C. violaceum.</h3>
 
                     <h3 class="sep30" style="display: none;">An inoculum of C. violaceum was left to make electrocompetent cells. Also, a test of antibiotic resistance was made in C. violaceum.</h3>
                     <h3 class="oct01" style="display: none;"> OD was measured 3 times in different times of the inoculum (every 45 minutes). Also, a miniprep was made trying a new kit.</h3>
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                     <h3 class="oct1" style="display: none;"> OD was measured 3 times in different times of the inoculum (every 45 minutes). Also, a miniprep was made trying a new kit.</h3>
                     <h3 class="oct02" style="display: none;">Digestions were made of the miniprep of the previous day. A growth kinetics curve was made from 9:20 to 18:50. Also we made electroporation of C. violaceum with RFP.</h3>
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                     <h3 class="oct2" style="display: none;">Digestions were made of the miniprep of the previous day. A growth kinetics curve was made from 9:20 to 18:50. Also we made electroporation of C. violaceum with RFP.</h3>
                     <h3 class="oct03" style="display: none;">We made electroporation of C. violaceum. Also, transformations were made of RFP (20F) in E. coli culture media LB+CAM.</h3>
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                     <h3 class="oct3" style="display: none;">We made electroporation of C. violaceum. Also, transformations were made of RFP (20F) in E. coli culture media LB+CAM.</h3>
                     <h3 class="oct04" style="display: none;">Inoculums were left to grow of the following: <br> Inducible promoter <br> GolS <br> Proton Pump <br> Sulfur Reductase <br> Iron Oxidase <br> Cyanide Hydratase <br> Inverter <br> G+R+T <br> Also, a purification was made of an electrophoresis gel of the fragments that contained pSB1C3. Inoculums were left of the promoter PR5 and Inverter. <br> Digestions were made of Sulfur reductase, GolS and Cyanide hydratase, afterwards an electrophoresis gel of these digestions. </h3>
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                     <h3 class="oct4" style="display: none;">Inoculums were left to grow of the following: <br> Inducible promoter <br> GolS <br> Proton Pump <br> Sulfur Reductase <br> Iron Oxidase <br> Cyanide Hydratase <br> Inverter <br> G+R+T <br> Also, a purification was made of an electrophoresis gel of the fragments that contained pSB1C3. Inoculums were left of the promoter PR5 and Inverter. <br> Digestions were made of Sulfur reductase, GolS and Cyanide hydratase, afterwards an electrophoresis gel of these digestions. </h3>
                     <h3 class="oct05" style="display: none;">Transformations were made of the following: <br>mRFP+2T= 20F <br>Inverter <br> Inducible Promoter <br> PR5 <br> Proton Pump <br> GolS <br> HCN <br> Sulfur Reductase <br> Iron Oxidase <br> Cyanide Hydratase <br> Minipreps were made of PR5, Inverter, Proton Pump.<br> Digestions were made of these Minipreps. <br> A purification was made of the electrophoresis gel that was made the day before of GolS, Cyanide Hydratase and Sulfur reductase. <br> Ligation of Sulfure reductase + pSB1C3+ GolS and Cyanide hydratase+pSB1C3 <br> Calcium competent C. violaceum were made based on the following spectrophotometric readings: <br> OD: 0.400 ** <br> OD: 0.500 * <br> OD: 0.600 ** </h3>
+
                     <h3 class="oct5" style="display: none;">Transformations were made of the following: <br>mRFP+2T= 20F <br>Inverter <br> Inducible Promoter <br> PR5 <br> Proton Pump <br> GolS <br> HCN <br> Sulfur Reductase <br> Iron Oxidase <br> Cyanide Hydratase <br> Minipreps were made of PR5, Inverter, Proton Pump.<br> Digestions were made of these Minipreps. <br> A purification was made of the electrophoresis gel that was made the day before of GolS, Cyanide Hydratase and Sulfur reductase. <br> Ligation of Sulfure reductase + pSB1C3+ GolS and Cyanide hydratase+pSB1C3 <br> Calcium competent C. violaceum were made based on the following spectrophotometric readings: <br> OD: 0.400 ** <br> OD: 0.500 * <br> OD: 0.600 ** </h3>
                     <h3 class="oct06" style="display: none;">Transformations were made of the part BBa_E0840 (RBS+GFP+2T). <br> Digestions were made from the following minipreps: PR5, Proton Pump, Inverter, and an TFP coding sequence for its backbone. <br> For later testing of the Proton Pump, the pH of LB medium was changed into 15 different tubes, into groups of 3 with pH 7, 8, 9, 10 and 11. <br> An electrophoresis Gel was run with today’s digestions, but the PR5 didn’t showed the desired results. Then, the parts were purified using a Aligen purification kit. <br> Afterwards, ligations of these parts was made, joining the proton pump with the backbone (psb1c3) with a 1:1 vector insert ratio. <br> Transformation of C. violaceum was made with RFP using Heat Shock (60 and 80 seconds) <br>0.1 μL of DNA and 50 μL of CaCl2 were used. <br> OD 0.500 cells and 60 seconds Heat Shock produced the largest number of transformed colonies.</h3>
+
                     <h3 class="oct6" style="display: none;">Transformations were made of the part BBa_E0840 (RBS+GFP+2T). <br> Digestions were made from the following minipreps: PR5, Proton Pump, Inverter, and an TFP coding sequence for its backbone. <br> For later testing of the Proton Pump, the pH of LB medium was changed into 15 different tubes, into groups of 3 with pH 7, 8, 9, 10 and 11. <br> An electrophoresis Gel was run with today’s digestions, but the PR5 didn’t showed the desired results. Then, the parts were purified using a Aligen purification kit. <br> Afterwards, ligations of these parts was made, joining the proton pump with the backbone (psb1c3) with a 1:1 vector insert ratio. <br> Transformation of C. violaceum was made with RFP using Heat Shock (60 and 80 seconds) <br>0.1 μL of DNA and 50 μL of CaCl2 were used. <br> OD 0.500 cells and 60 seconds Heat Shock produced the largest number of transformed colonies.</h3>
                     <h3 class="oct07" style="display: none;">Transformations of 3 different promoters and an RFP without promoter were made: <br> 5C K608004 (PR2) Promoter <br> 5E K608006 (PR5) <br> 3O K608002 <br> 20 F (RFP) <br> Digestions were also made of the Inducible promoter and the PR5. <br> Transformation of the ligations of G,S, C also of the PR5 promoter and the 4 parts previously stated. <br> Gel extraction of the Heat Shock Promoter. <br> Calcium competent cells were made of BL21.</h3>
+
                     <h3 class="oct7" style="display: none;">Transformations of 3 different promoters and an RFP without promoter were made: <br> 5C K608004 (PR2) Promoter <br> 5E K608006 (PR5) <br> 3O K608002 <br> 20 F (RFP) <br> Digestions were also made of the Inducible promoter and the PR5. <br> Transformation of the ligations of G,S, C also of the PR5 promoter and the 4 parts previously stated. <br> Gel extraction of the Heat Shock Promoter. <br> Calcium competent cells were made of BL21.</h3>
                     <h3 class="oct08" style="display: none;">Transformations of the BL21 cells were made with Gol S (G), Cyanide hydratase(C) and Sulfur reductase(S), all with resistance to kanamycin. </h3>
+
                     <h3 class="oct8" style="display: none;">Transformations of the BL21 cells were made with Gol S (G), Cyanide hydratase(C) and Sulfur reductase(S), all with resistance to kanamycin. </h3>
                     <h3 class="oct09" style="display: none;">The following DNA was gathered from the iGEM plate 3 of 2014: <br> BBa_J06702 → CAMr. RBS + RFP + 2T (280 bp). Total size: 2940 bp. <br> BBa_K608003 → CAMr. Promoter + RBS (56 bp). Total size: 2100 bp. <br> BBa_K608006 → CAMr. Promotor + RBS (56 bp). Total size: 2100 <br>Transformation of BL21 calcium competent cells from October 7 using the same conditions as the ones used on October 8, with BBa_J06702 (1), BBa_K608003 (2), BBa_K608006 (3), and RFP (4). </h3>
+
                     <h3 class="oct9" style="display: none;">The following DNA was gathered from the iGEM plate 3 of 2014: <br> BBa_J06702 → CAMr. RBS + RFP + 2T (280 bp). Total size: 2940 bp. <br> BBa_K608003 → CAMr. Promoter + RBS (56 bp). Total size: 2100 bp. <br> BBa_K608006 → CAMr. Promotor + RBS (56 bp). Total size: 2100 <br>Transformation of BL21 calcium competent cells from October 7 using the same conditions as the ones used on October 8, with BBa_J06702 (1), BBa_K608003 (2), BBa_K608006 (3), and RFP (4). </h3>
 
                     <h3 class="oct10" style="display: none;">Single colonies were isolated from cultures (1) and (4) from October 9 and inoculated with kanamycin at 6:50 pm. <br> Calcium competent BL21 cells were transformed via Heat Shock (60 seconds), using 1μL DNA: <br> Part K516030 (Kit plate 1_2015, 9I) → mRFP protein generator (w/o promotor) <br> Part K823006 (Kit plate 2_2015, 20M) → Anderson promoter S23102 <br> Part K823008 (Kit plate 1_2015, 22A) → Anderson promoter S23106 <br> Minipreps were made of PR2 and PR5 with K608004/K608006, promotor and RBS. <br> Once digestions were made with SP, 20 μL final volume, an electrophoresis gel was run using said digestion products.</h3>
 
                     <h3 class="oct10" style="display: none;">Single colonies were isolated from cultures (1) and (4) from October 9 and inoculated with kanamycin at 6:50 pm. <br> Calcium competent BL21 cells were transformed via Heat Shock (60 seconds), using 1μL DNA: <br> Part K516030 (Kit plate 1_2015, 9I) → mRFP protein generator (w/o promotor) <br> Part K823006 (Kit plate 2_2015, 20M) → Anderson promoter S23102 <br> Part K823008 (Kit plate 1_2015, 22A) → Anderson promoter S23106 <br> Minipreps were made of PR2 and PR5 with K608004/K608006, promotor and RBS. <br> Once digestions were made with SP, 20 μL final volume, an electrophoresis gel was run using said digestion products.</h3>
 
                     <h3 class="oct11" style="display: none;">C. violaceum transformed with RFP and resistance to kanamycin was cultured on solid medium. <br> A stock of C. violaceum + RFP was made for cryopreservation at -80°C with 20% glycerol. <br> A stock of E. coli BL21 transformed with GolS and resistance to ampicilyn was made for cryopreservation at -80°C with 20% glycerol. </h3>
 
                     <h3 class="oct11" style="display: none;">C. violaceum transformed with RFP and resistance to kanamycin was cultured on solid medium. <br> A stock of C. violaceum + RFP was made for cryopreservation at -80°C with 20% glycerol. <br> A stock of E. coli BL21 transformed with GolS and resistance to ampicilyn was made for cryopreservation at -80°C with 20% glycerol. </h3>
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Latest revision as of 19:22, 2 December 2016

iGEM 2016 - Tec de Monterrey




July

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3 4 5 6 7 8 9
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17 18 19 20 21 22 23
24 25 26 27 28 29 30
31 1 2 3 4 5 5

August

30 1 2 3 4 5 6
7 8 9 10 11 12 13
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21 22 23 24 25 26 27
28 29 30 31 1 2 3
4 5 6 7 8 9 10

September

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4 5 6 7 8 9 10
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18 19 20 21 22 23 24
25 26 27 28 29 30 1
2 3 4 5 6 7 8

October

25 26 27 28 29 30 1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31 1 2 3 4 5

Thursday June 30, 2016

Saturday July 2, 2016

Monday July 4, 2016

Tuesday July 5, 2016

Wednesday July 6, 2016

Thursday July 7, 2016

Friday July 15, 2016

Saturday July 16, 2016

Monday July 18, 2016

Tuesday July 19, 2016

Wednesday July 20, 2016

Thursday July 21, 2016

Friday July 22, 2016

Saturday July 23, 2016

Tuesday July 26, 2016

Wednesday July 27, 2016

Thursday July 28, 2016

Friday July 29, 2016

Monday August 1, 2016

Tuesday August 2, 2016

Wednesday August 3, 2016

Thursday August 4, 2016

Friday August 5, 2016

Tuesday August 9, 2016

Thursday August 11, 2016

Friday August 12, 2016

Saturday August 13, 2016

Monday August 15, 2016

Tuesday August 16, 2016

Wednesday August 17, 2016

Thursday August 18, 2016

Friday August 19, 2016

Saturday August 20, 2016

Sunday August 21, 2016

Monday August 22, 2016

Tuesday August 23, 2016

Wednesday August 24, 2016

Thursday August 25, 2016

Saturday August 27, 2016

Monday August 29, 2016

Tuesday August 30, 2016

Wednesday August 31, 2016

Thursday September 1, 2016

Friday September 2, 2016

Saturday September 3, 2016

Sunday September 4, 2016

Monday September 5, 2016

Friday September 9, 2016

Saturday September 10, 2016

Monday September 12, 2016

Tuesday September 13, 2016

Wednesday September 14, 2016

Friday September 16, 2016

Saturday September 17, 2016

Sunday September 18, 2016

Monday September 19, 2016

Tuesday September 20, 2016

Wednesday September 21, 2016

Thursday September 22, 2016

Tuesday, September 27, 2016

Friday, September 30, 2016

Saturday, October 01, 2016

Sunday, October 02, 2016

Monday, October 03, 2016

Tuesday, October 04, 2016

Wednesday, October 05, 2016

Thursday, October 06, 2016

Friday, October 07, 2016

Saturday, October 08, 2016

Sunday, October 09, 2016

Monday, October 10, 2016

Tuesday, October 11, 2016

Wednesday, October 12, 2016

Thursday, October 13, 2016

Friday, October 14, 2016

Saturday, October 15, 2016

Sunday, October 16, 2016

Monday, October 17, 2016

Wednesday, October 19, 2016




We tested our antibiotics: ampicillin, kanamycin, chloramphenicol with our PLYS and Shuffle bacteria





Brought to you by iGEM Tec-Monterrey 2016