Team:USP-UNESP-Brazil/Notebook

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===First assemblies confirmed===
===First assemblies confirmed===
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After this messy period trying to fix the assembly's problems, the  good results came. We made with fully success QC1, QC4, QC5, QR4, QR5 and QR1 assemblies through 3A protocol. The results and Agarose Gel photos follows.  
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After this messy period trying to fix the assembly's problems, the  good results came. We made with fully success QC1, QC4, QC5, QR4, QR5, QR6 and QR1 assemblies through 3A protocol. The results and Agarose Gel photos follows.  
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16/06/2012 - Otto, Digestion
16/06/2012 - Otto, Digestion
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17/06/2012 - Daniel, Electrophoresis Gel
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17/06/2012 - Daniel, Electrophoresis Gel([https://static.igem.org/mediawiki/2012/c/ce/QC1_e_QC6_-_2.png QC1 and QC6 Electrophoresis Gel])
====QC4====
====QC4====
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13/06/2012 - Pedro, Digestion
13/06/2012 - Pedro, Digestion
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17/06/2012 - Pedro, Electrophoresis Gel
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17/06/2012 - Pedro, Electrophoresis Gel([https://static.igem.org/mediawiki/2012/a/a7/QC4_120813.png QC4 Electrophoresis Gel])
====QC5====  
====QC5====  
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18/08/2012 - Daniel, Electrophoresis Gel
18/08/2012 - Daniel, Electrophoresis Gel
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([https://static.igem.org/mediawiki/2012/b/b0/QC5_-_120818.png QC5 Electrophoresis Gel])
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15/06/12 – Fernando: All colonies were still red, showing that the assembly did not work.
15/06/12 – Fernando: All colonies were still red, showing that the assembly did not work.
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([https://static.igem.org/mediawiki/2012/3/32/QR4.png QR4 Electrophoresis Gel])
====QR5====
====QR5====
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07/08/12 – Fernando: Each inoculate was used for DNA extraction. 3uL plasmid from each of the four colonies were digested with EcoRI and PstI enzymes. QR5 assembly was confirmed with gel electrophoresis (three out of the 4 colonies were confirmed).
07/08/12 – Fernando: Each inoculate was used for DNA extraction. 3uL plasmid from each of the four colonies were digested with EcoRI and PstI enzymes. QR5 assembly was confirmed with gel electrophoresis (three out of the 4 colonies were confirmed).
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([https://static.igem.org/mediawiki/2012/2/29/QR5.png QR5 Electrophoresis Gel])
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====QR6====
====QR6====
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20/09/12 – Fernando: Since 3A Assembly Protocol was not working to assemble the QR1 construct, each part (12H, 12M and pSB1C3) were digested using the 3A Assembly Protocol digestion procedure and the total volume was loaded in a electrophoresis gel. The idea was to purify the inserts and linearized plasmid backbone pSB1C3 using Quiagen Gel Extraction Kit. Since the bands could barely be seen under U.V, nothing was purified.
20/09/12 – Fernando: Since 3A Assembly Protocol was not working to assemble the QR1 construct, each part (12H, 12M and pSB1C3) were digested using the 3A Assembly Protocol digestion procedure and the total volume was loaded in a electrophoresis gel. The idea was to purify the inserts and linearized plasmid backbone pSB1C3 using Quiagen Gel Extraction Kit. Since the bands could barely be seen under U.V, nothing was purified.
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([https://static.igem.org/mediawiki/2012/7/7f/QR6.jpg QR6 Electrophoresis Gel])
====QR1====
====QR1====
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21/09/2012 - Pedro - Digestion and PCR.
21/09/2012 - Pedro - Digestion and PCR.
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24/09/2012 - Pedro - Electrophoresis Gel.
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24/09/2012 - Pedro - Electrophoresis Gel.([https://static.igem.org/mediawiki/2012/9/91/QR1_120924b.png QR1 Electrophoresis Gel])

Latest revision as of 03:54, 27 September 2012