Team:Brasil-USP/Notebook/Calendar

Notebook

Calendar

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)


    		 16/06
    - 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

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