Protocols & Notebook

Protocols

  • Bacillus Subtilis Transformation {DF}
  • Conoly PCR for E. coli PDF
  • Chemical transformation for E. coli PDF
  • E. coli Heat-shock transformation PDF
  • Golden Gate Assembly using the Echo liquid handler
  • Making electrocompetent E. coli* cells PDF
  • Gel Electrophoresis PDF
  • Gibson Assembly PDF
  • Glycerol stocks PDF
  • Golden Gate Assembly PDF
  • Home-made E. coli competent cells with CaCl2. PDF
  • Sequencing PDF
  • E. coli transformation with TSS PDF

Lab Notebook

July 15, 2022 :

    - Transformation pETDuet- p15

Jul 17, 2022 :

    - Liquid cultures of pACYCDuet-1 p17/pCDFDuet-1 p16/pCOLADuet-1 p18

Jul 18, 2022 :

    - Antibiotic Resistance Assay - Miniprep : pACYCDuet-1 p17/pCDFDuet-1 p16/pCOLADuet-1 p18 - Transformation dpB.015, dpB016, ppB.030 - Tranformation pETDuet-p15

Jul 19, 2022:

    - Miniprep : pACYCDuet-1 p17/pCDFDuet-1 p16/pCOLADuet-1 p18 - Transformation :ppB.030/ppb.029 ; ppB.028/ppB.029 ; ppB.028/29/30 - Transformation: ppB.030/ppB.0.28

Jul 20, 2022:

    - Transformation of p15 in commercial comp cells - Liquid culture all pDuet( exept pET) + GFP antoine + Moclo DVA et DVE

Jul 26, 2022:

    - CIDAR MoClo Vector UPGRADE - IO rodeo potentiostat, electrode wiring - Cloning PnasA for B.Subtilis

Jul 28, 2022:

    - Cloning Lvl1

Aug 1, 2022:

    - Miniprep: dpB 004, 006, 010, 013, 016, 075 - NanoDrop of minipreps - First test of potentiostat (success!) - Cultures on kanamycin plates

Aug 2, 2022 :

    - UV view on cultures - Miniprep (NEW PROTOCOL): dpB 006, pp038 - NanoDrop miniprep dpB006, pp038 - Cloning of dp.012 - Colony PCR + gel v35

Aug 3, 2022 :

    - Glycerol stock - Colony PCR v35 + gel

Aug 4, 2022 :

    - Transformation dpB.008 dpB.009 dpB.011 - Miniprep by new protocol 1st part, nanodrop, liquid culture - Colony PCR

Aug 7, 2022 :

    - Miniprep by new protocol 2nd part + results analysis

Aug 9, 2022:

    - Sequencing dpB.003,dpB.005,dpB.006,dpB.007,dpB.008, dpB 009, dpB.011, dpB.012, dpB.013

Aug 10, 2022:

    - Cloning for dpB 018 & 019 (Act106 & Inv106) from JL - Miniprep dpB.001, dpB.002, dpB.006, dpB.010, dpB.014

Aug 12, 2022:

    - Miniprep dpB 018 & 019 (Act 106 & Inv 106)

Aug 14,2022:

    - Miniprep dpB 018 & 019 (Act 106 & Inv 106)

Aug 15, 2022:

    - Processor parts building and characterisation - Test of diff.concentration of inducers for dpB.010

Aug 16, 2022:

- Testing Microscope slides for Hardware

Aug 17, 2022:

    - Simple OD Meter - Miniprep Moclo parts for Echo (pp.048, pp.049,pp.050)

Aug 18, 2022:

    - Upgade pDuet with V35, Vector cloning - Primer dilution and sequencing prep

Aug 19, 2022:

    - Plate Reader Inducers Marionettes

Aug 21, 2022:

    - Golden Gate pDuet and G Blocks - Bacteria Electroshock

Aug 22, 2022:

    - Plate Reader Assay Inv & Act - Transformation of b.sub - Perform lacZ screening assay - Perfom starch test - Characterisation of the promotor - Building all pDuet with the g.block and try different combinaison to test the processor and the marionettes system with multiple plasmids

Aug 23, 2022:

    - building of all dpB in the 3 pDUET by goldengate - Plate Reader Assay pOxyS - Get the electroshock 96 well plate as well as the microscope slide ready for experiments - Ideally get one experiment on Amplitude running - Get the Uri Alon Library working for experiments (prepare batch culture and clone the library)

Aug 24, 2022:

    - Transformation of all pDUET in comp cells

Aug 25, 2022:

    - Plate Reader Assay pSoxR

Aug 26,2022:

    - Plate Reader assay pOxyS - Higher concentrations

Aug 27, 2022:

    - Microscope electroshock - Subtitoolkit Replica Plate - Electrical conductivity meter - 555 Timer

Aug 28,2022 :

    - Gibson Assembly Debug - Sequencing pDuets

Aug 29, 2022:

    - Microscope electroshock - Uri Alon Library

Sep 1, 2022:

    - Lac Z colony PCR

Sep 2, 2022:

    - Output Conductivity Characterization #1

Sep 8, 2022:

    - [FACS] Electroshocking bacteria on 96 Well plate

Sep 10, 2022

    - Miniprep for Paris_Inv Gibson - Growth Curve Uri Alon m9

Sep 11, 2022

    - PCR for Paris_Inv

Sep 13, 2022:

    - PCR for Paris_Inv Q5

Sep 15, 2022:

    - Transformation Partnership Training Constructs

Sep 18, 2022:

    - Streak of Paris_Inv and Training constructs - Gibson Assembly Paris_Inv - PCR Clean-up

Sep 20, 2022:

    - Colony PCR G3-1 and 7 G6 + gel - [FACS] Electroshocking bacteria on 96 Well plate

Sep 21, 2022:

    - Miniprep of STK parts for Echo and Gibson Paris_Inv for seq - [FACS] Electroshocking bacteria on 96 Well plate

Sep 22, 2022:

    - Echo prep STK

Sep 27, 2022:

    - Chronoamperometry Paris_Inv - 1 hour

Sep 28, 2022:

    - STK 110 plating - Chronoamperometry Paris_Inv - 8 min

Sep 29, 2022:

    - Chronoamperometry - 6 hours and 12 hours - Pduet Plate reader experiment

Sep 30, 2022:

    - Chronoamperometry pOxyS - 6 hours - Chart characterization

Oct 2, 2022:

    - Plate Reader Output Experiment - Activation Inverter PsoxR

Oct 6, 2022:

    - Chronoamperometry - 16 hours