July

Monday	Tuesday	Wednesday	Thursday	Friday	Saturday	Sunday
		01/06 - Agarose gel (RBS_hokD) - Gel purification (RBS_hokD) - Ligation 2X_RBS_hokD (RBS_hokD + RBS_hokD) - EXSP Digestion (P0040, Pl_RBS, GFP_R0440) - Agarose gel (P0040) - Gel purification (P0040) - Ligation Pt_I13507(P0040 + I13507) - Interlab gel purification and ligation 02/06 - Transformation Ptet_I13507 - Agarose gel (Pl_RBS, GFP_P0440) - Gel purification (Pl_RBS, GFP_P0440) - Ligation Pl_P0440 (Pl_RBS + GFP_P0440) - Interlab transformation (with wrong antibiotic ampicillin - is possible found in registry the same parts in another backbone BBa_J61002)	03/06 - Transformation (Pr_RBS, 2X_RBS_hokD) - EXSP digestion (P0440, R0040) - Transformation Pl_R0440 (Pl_RBS + GFP_P0440) - Inoculum (Pt_I13507)	04/06 - Agarose gel (P0440, R0040) - Gel purification (P0440, R0040) - Ligation tet_Ptet (P0440 + R0040) - Inoculum (2X_RBS_hokD) - Miniprep (Pt_I13507)	05/06 - Inoculum (R0080, K91403) - EXSP digestion (tet_Term, Pt_E0840) - Miniprep (2X_RBS_hokD)
06/06 - NotI digestion (2X_RBS_hokD, Pr_RBS, Pa_RBS) - Transformation (tet_Ptet) - Agarose gel (2X_RBS_hokD) – positive results - Agarose gel (tet_Term, Pt_E0840) - EXSP digestion (Pt_E0480) - Inoculum (Pl_P0440) - NotI digestion (Pt_I13507) – positive results	07/06 - Agarose gel (Pr_RBS, Pa_RBS, Pt_E0480) – positive results for Pr_RBS and Pt_E0840) - EXSP digestion (2X_RBS_hokD, B0015) - Ligation Term_E0840 (Pt_E0840 + tet_Term) - Inoculum (tet_Ptet) - Transformation (Term_E0840) - Miniprep (Pl_P0440) - Not digestion (Pl_P0440) – positive results - EXSP digestion (Pl_P0440, Pt_I13507) - Inoculum (GFP_P0440)	08/06 - Agarose gel (2X_RBS_hokD, B0015) – B0015 didn’t digest - Gel purification (2X_RBS_hokD) - EXSP digestion (B0015) - Miniprep (tet_Ptet) - NotI digestion (tet_Ptet) - Agarose gel (tet_Ptet, B0015) – positive results - Gel digestion (B0015) - Ligation hokD_Term (2X_RBS_hokD + B0015) - Ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - Miniprep (GFP_P0440) - EXSP digestion (GFP_P0440)	09/06 - Agarose gel and gel purification (GFP_P0440) - Ligation GFP_I13507 (GFP_P0440 + Pt_I13507) - Transformation (Pl_I13507)	10/06 - Transformation (hokD_Term) - Inoculum (Pl_I13507) - Transformation (GFP_I13507)	11/06 - Miniprep (Pl_I13507)	12/06
13/06 - Inoculum (GFP_I13507) - NotI digestion (Pl_I13507) - Agarose gel (Pl_I13507) – negative results	14/06 - Miniprep (GFP_I13507) - NotI digestion (GFP_I13507) – negative results - EXSP digestion (Pl_P0440, Pt_I13507) - Inoculum (GFP_P0440, Pt_I13507)	15/06 - Agarose gel (Pl_P0440, Pt_I13507) - Gel purification (Pl_P0440, Pt_I13507) - Ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - Miniprep (GFP_P0440, Pt_I13507) - EXSP digestion (GFP_P0440, Pt_I13507)	16/06 - Agarose gel and gel purification (GFP_P0440, Pt_I13507) - Ligation GFP_I13507 (GFP_P0440 + Pt_I13507) - Transformation (Pl_I13507)	17/06 - Transformation (GFP_I13507) - Inoculum (Pt_Term, Pl_I1350)	18/06 - Miniprep (Pt_Term, Pl_I13507) - NotI digestion (Pt_Term, Pl_I13507) - Agarose gel (Pt_Term, Pl_I13507) – positive results for Pt_Term	19/06
20/06 iGEM kit arrived!	21/06 - Inoculum (GFP_I13507, Pl_I13507) - New interlab transformations	22/06 - Miniprep (GFP_I13507, Pl_I13507) - NotI digestion (GFP_I13507, Pl_I13507) - EXSP digestion (Pr_RBS, R0080) - Agarose gel (GFP_I13507, Pl_I13507, Pr_RBS) - Gel purification (Pr_RBS) - Inoculum (GFP_I13507, Pl_I13507) - Interlab inocula	23/06 - Agarose gel (R0080) - Miniprep (GFP_I13507, Pl_I13507) - NotI digestion (GFP_I13507, Pl_I13507) - Agarose gel (GFP_I13507, Pl_I13507) – Positive results for GFP_I13507 - EXSP digestion (GFP_I13507) - Interlab miniprep and restriction analysis	24/06 - Miniprep (B0015, Pt_P0440) - EXSP digestion (2X_RBS_hokD, B0015) - Transformation (Pa_RBS) - Agarose gel of restriction analysis: devices confirmed	25/06 - agarose gel (2X_RBS_hokD, B0015) - B0015 did not digest	26/06 - Agarose gel and gel purification (GFP_I13507) - Ligation Pr_I13507 (Pr_RBS + GFP_I13507)
27/06 - EXSP digestion (Pl_RBS, Pt_Term) - Transformation (Pr_I13507, Pa_RBS)	28/06 - Agarose gel (Pl_RBS, Pt_Term) ligation Pr_I13507(Pr_RBS + Pt_Term)	29/06 - EXSP digestion (B0015, 2X_RBS_hokD) - agarose gel (B0015, 2X_RBS_hokD) - 2X_RBS_hokD didn’t digest - EXSP digestion overnight (2X_RBS_hokD) - transformation (Pr_Term, Pl_RBS)	30/06 - inoculum (Pl_RBS, Pa_RBS) - agarose gel (B0015, 2X_RBS_hokD) - Digestion worked! - ligation 2X_hokD_Term (B0015 + 2X_RBS_hokD) P0440_hokD(P0440_term + 2X_RBS_hokD)	31/06

August

Monday	Tuesday	Wednesday	Thursday	Friday	Saturday	Sunday
					01/08	02/08
03/08 - EXSP digestion (R0010, R0080, B0034, Pt_I13507) - NotI digestion (Pl_I13507) - Ligation Pa_RBS (R0080 + B0034) and Pl_RBS (R0010 + B0034)	04/08 - NotI digestion (Pr_I13507) - Agarose gel (Pr_I13507) - Positive results!! - EXSP digestion (Pt_I13507) - Transformation (Pl_RBS and Pa_RBS)	05/08 - agarose gel (Pl_I13507, Pt_I13507)- negative results for Pl_13507 :( - Agarose gel and gel purification (Pt_I13507) - ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - inoculum (Pl_RBS, Pa_RBS)	06/08 - Transformation (Pl_I13507, Pr_I13507) - miniprep (Pl_RBS, Pa_RBS) - NotI digestion (Pl_RBS, Pa_RBS)	07/08 - Agarose gel (Pl_RBS, Pa_RBS) - Only Pa_RBS was positive - EXSP digestion (Pa_RBS, Pt_Term) - Ligation Pa_Term(Pa_RBS + Pt_Term) Pa_I13507 (Pa_RBS + GFP_I13507)	08/08 - Ligation (2X_RBS_hokD + Term) - Transformation (Pa_I13507, Pa_Term)	09/06 - Inoculum (Pa_I13507,Pl_I13507,Pa_Term)
10/08 - miniprep (Pa_I13507, Pl_I13507, Pa_Term) - NotI digestion (Pa_I13507, Pa_Term) - Agarose gel (Pa_I13507, Pl_I13507, Pa_Term, Pl_RBS) - Positive results for Pa_I13507 and Pa_Term - Pre-inoculum of interlab study devices, BBa_K876004, and controls	11/08 - Vector exchange digestion (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - Ligation with pSB1C3 (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - EXSP digestion (Pl_RBS, GFP_I13507, Pt_Term) - ligation (Pl_I13507, Pl_Term) - Transformation OmpA - Sample preparation of Interlab Study devices and controls (plate reader) - End-point measurement of Interlab Study devices and controls - Sample preparation of BBa_K876004 part (plate reader) - Kinetic measurement of BBa_K876004 (14 hours).	12/08 - Transformation (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - Inoculum OmpA - Pre-inoculum of interlab study devices, BBa_K876004, and controls	13/08 - Inoculum (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - miniprep OmpA - EXSP digestion (B0034, OmpA) - agarose gel (B0034, OmpA) - Ligation RBS_OmpA (RBS + OmpA) - Sample preparation of Interlab Study devices and controls (flow cytometry) - Measurement of Interlab Study devices and controls - Sample preparation of BBa_K876004 part (flow cytometry) - Flow cytometry measurement of BBa_K876004 (14 hours).	14/08 - miniprep (Pr_RBS, GFP_P0440(C3), Pl_I13507) - Positive results for Pl_I13507 - and GFP_P0440 - Transformation RBS_OmpA	15/08 - Inoculum (RBS_OmpA, GFP_I13507(C3)	16/08 - miniprep (RBS_OmpA, GFP_I13507(C3)) - NotI digestion (RBS_OmpA, GFP_I13507(C3)) - Agarose gel (RBS_OmpA, GFP_I13507(C3)) - Ligation Pl_Term(Pl_RBS + Pl_Term)
17/08 - Inoculum (RBS_OmpA) - Transformation (Pl_Term)	18/08 - IDT primers and gBlocks arrived - miniprep (RBS_OmpA) - NotI digestion (RBS_OmpA) - Agarose gel (RBS_OmpA) - Postive - Vector exchange digestion (RBS_hokD, Pr_RBS) - Ligation RBS_hokD(C3), Pr_RBS(C3) - Inoculum (Pl_Term)	19/08 - Transformation (RBS_hokD(C3), Pr_RBS(C3)) - NotI digestion (Pl_Term) - Agarose gel (Pl_Term)- positive!!!	20/08 - gBlocks and Primers resuspension and quantification - EP digestion (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term) - Ligation Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term with pBS1C3 - Inoculum (RBS_hokD(C3), Pr_RBS(C3))	21/08 - PCR for mutation of EcoRI roxA site - Agarose gel for amplification confirmation and purification of roxA PCR mutation - DpnI digestion of purificated PCR and DH5a transformation using the DNA digested - PCR for insert amplification of Lcp inserting Biobrick sites - PCR for insert amplification of Lcp inserting pETSUMO restriction sites (Lcp heterologous expression) - Agarose gel for amplification confirmation and purification of lcp PCRs - Digestion of Lcp PCR biobrick and pSB1C3 with EP - Digestion of Lcp PCR for pETSUMO and pETSUMO with restriction enzymes BamHI and HindIII - miniprep (RBS_hokD(C3), Pr_RBS(C3)) - NotI digestion (RBS_hokD(C3), Pr_RBS(C3)) - Agarose gel (RBS_hokD(C3), Pr_RBS(C3)) - All negative - Transformation (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term) - Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls)	22/08 - Agarose gel purification for Lcp ligations - Transformation of Lcp for biobrick and Lcp for pETSUMO using insert and vector purified - Inoculum of 6 roxA mutant colonies - Inoculum (Pl_I13507(C3)) - Sample preparation (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls) (plate reader) - five 96-well plates - Measurement of Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls (12 hours - plate reader) - Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (codes))	23/08 - gBlock reactions and transformations (linker-GFP, 6XHis, signalTAT) - Miniprep roxA mutants - Digestion of EcoRI and HindIII of roxA mutants and plasmid non-mutated (control) - Agarose gel of the restriction analysis - Sample preparation (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (plate reader) - five 96-well plates - Measurement of Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507 and BBa_K876004 (12 hours - plate reader) - miniprep (Pl_I13507(C3)) - NotI digestion (Pl_I13507(C3)) - Agarose gel (Pl_I13507(C3)) - posite results
24/08 - Inoculum (linker-GFP, 6XHis, signalTAT, Lcp(pETSUMO), Lcp(pSB1C3)) - miniprep (Pa_Term, Pl_Term) - EP digestion (Pa_Term, Pr_Term, Pl_Term, 2xHokD, B0015) - PCR of mutant roxA for biobrick ligation	25/08 - miniprep (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - NotI and EXSP digestions ( (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Agarose gel ((OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Positive results for OmpA_RBS ,linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3) - Ligation 6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3) - Ligation (Pa_Term(C3), Pl_Term(C3), Pr_Term(C3) - Agarose gel (mutant RoxA)	26/08 - Transformation 6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp	27/08 - Inoculum (6xHis_Term, GFP_Term, mpA_Lcp, Tat_Lcp) - Transformation (2xhokD_Term) - PCR of mutant roxA for biobrick ligation - Agarose gel of mutant roxA PCR product	28/08 - roxA PCR product purification - roxA PCR product digestion with EP - miniprep (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp) - NotI and EXSP digestions (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp) - Only 6xHis_Term was negative - ligation Lcp_Term I (Tat_Lcp + GFP_Term), Lcp_Term II (OmpA_Lcp + GFP_Term) - Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)	29/08 - Agarose gel (roxA PCR product) - Ligation roxA(mutant) +pSB1C3 - Sample preparation (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) (plate reader) - seven 96-well plates - Measurement of test promoters (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507)	30/08 - Transformation (roxA)
31/08 - Inoculum (roxA) - Transformation (Lcp_Term I, Lcp_Term II)

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