Difference between revisions of "Team:Brasil-USP/Notebook/Calendar"

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Revision as of 02:28, 18 September 2015

Calendar

Notebook

July

© 2015 Brasil-USP Team · iGEM · IFSC · USP

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Monday	Tuesday	Wednesday	Thursday	Friday	Saturday	Sunday
30/06 PCR lasR (BBa_C0079). The purpose of this PCR was to add the RBS (BBa_K143021), through addition of the sequence in the primer foward, and also to remove the LVA tag. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this part will be called BBa_K1521001.	01/07 Gel Purification of lasR PCR product	02/07 Transformation of the Biobricks: BBa_K143012 BBa_K081001 BBa_E0840 Inoculum of the Biobricks sent by iGEM HQ: BBa_K316037 BBa_K316018 BBa_K316015	03/07 Transformation of the Biobricks sent by the Imperial College Team: BBa_K316016 BBa_K143031 Miniprep, Quantification of the plasmidial DNA samples and Restriction Analysis: BBa_K316037 BBa_K316018 BBa_K316015	04/07	05/07	06/07 Inoculum: BBa_K143012 BBa_K081001 BBa_E0840
07/07 Miniprep, Quantification and Restriction Analysis: BBa_K143012 BBa_K081001 BBa_E0840 Transformation: BBa_B0015	08/07	09/07 Inoculum: BBa_B0015	10/07 Miniprep, Quantification and Restriction Analysis: BBa_B0015 (restriction analysis fail, repeat) PCR the qteE gene for amplification. We also added the RBS (BBa_K143021,) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called BBa_K1521000. Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)	11/07 Prepare 40% glycerol solution Transformation: RBS + lasR (BBa_C0079) Gel Purification: RBS + qteE	12/07	13/07
14/07 Assembly:AIII Digestion EXSP Prepare X-gal Prepare more LB medium (solid + liquid)	15/07 Assembly:AIII Quantification of digestion EXSP products Replate RBS+lasR Ligation of PCR qteE in pGEM T-easy vector Transformation: BBa_J04450 Restriction analysis: BBa_B0015	16/07 Transformation: BBa_K823003 RBS+qteE Electrophoresis analysis: BBa_B0015	17/07 Glycerol Stock and Miniprep: BBa_J04450 RBS+lasR BBa_B0015 Digestion: BBa_J04450 (EP) BBa_J04450 (XS) Inoculum: BBa_K823003 Transformation BBa_K143055	18/07 Glycerol Stock and Miniprep: BBa_J04450 BBa_B0015 Gel Purification: BBa_J04450 (EP) BBa_J04450 (XS) Restriction analysis: BBa_K823003	19/07	20/07
21/07 Transformation: BBa_K316016 Inoculum: BBa_K143055 RBS+qteE Assembly KI: Digestion EXSP	22/07 Inoculum: BBa_K316016 Miniprep, Restriction analysis and Digestion: RBS+qteE Assemblies AIII and KI: Gel Purification Ligation	23/07 Assemblies AIII and KI: Transformation Prepare more competent cells Prepare LB medium (solid and liquid) Inoculum BBa_K316016 Prepare RBS+lasR and RBS+qteE for sequencing	24/07 Assemblies AIII and KI: Inoculum Miniprep: BBa_K316016	25/07 Assemblies AIII and KI: Glycerol Stock Miniprep Restriction analysis	26/07	27/07
28/07 Assembly AIII: Restriction analysis (+NdeI enzyme) OBS: This analysis was reapeted because we had problems confirming the assembly. The issue was that both the construction and the pSB1C3 vector had similar size. Assembly AII, AV and AVI: Digestion EXSP	29/07 >Assembly AII, AV and AVI: Gel Purification Ligation	30/07 >Assembly AII, AV and AVI: Transformation	31/07 Assembly AII, AV and AVI: Inoculum PCR BBa_K143055 for RBS removal OBS: This was one of the parts sent by the Imperial College Team. Because of a comunication failure we assumed that this part was comming without the RBS, so we included a RBS in the parts we were going to use with this Lac promotor :) PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector