Team:NCTU Formosa/Notebook

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Notebook

July 1

Build UV model

July 2

Build mutation analytic program

July 3

None

July 4

None

July 5

None

July 6

None

July 7

None

July 8

None

July 9

None

July 10

None

July 11

Fix mutation analytic program

July 12

None

July 13

None

July 14

Fix hypermutation model

July 15

IDT gblock resuspend

IDT Taq PCR

  • Sample:All IDT parts.

July 16

Dilute IDT solutions

  • Sample:
    • hipA, LuxI-LuxR(without Pcon), LuxI-LuxR, GFP-Ampicillin.
      (Time set: 95°C/5min, [95°C/30sec, 55°C/30sec, 72°C/143sec]x25cycles, 72°C/10min)
    • ccdB, MazF, yafQ, ydfD, chpBK, hipA.
      (Time set: 95°C/5min, [95°C/30sec, 55°C/30sec, 72°C/91sec]x25cycles, 72°C/10min)

IDT dilution

  • Dilute the gblock solution 50-fold

Taq PCR & gel electrophoresis

  • Sample:
    • hipA, LuxI-LuxR(without Pcon), LuxI-LuxR, GFP-Ampicillin
      (Time set: 95°C5/min, [95°C/30sec, 55°C/30sec, 72°C/143sec]x35cycles, 72°C/10min)
    • ccdB, MazF, yafQ, ydfD, chpBK, hipA.
      (Time set: 95°C/5min, [95°C/30sec, 55°C/30sec, 72°C/91sec]x35cycles, 72°C/10min)
    • ccdB, yafQ.
      (Time set: 95°C/5min, [95°C/30sec, 48°C/30sec, 72°C/75sec]x35cycles, 72°C/10min)
    • LuxI-LuxR(without Pcon).
      (Time set: 95°C/5min, [95°C/30sec 48°C/30sec, 72°C/143sec]x35cycles, 72°C/10min)

July 17

Taq PCR & gel electrophoresis

  • Sample:LuxI-LuxR(without Pcon)

Pfu PCR & gel electrophoresis

  • Sample:ccdB, yafQ

Gel extraction

  • Sample:ccdB, yafQ

July 18

Pfu PCR & gel electrophoresis

  • Sample:ccdB, yafQ

Gel extraction

  • Sample:ccdB, yafQ.

July 19

Pfu PCR & gel electrophoresis

  • Sample:ccdB, yafQ

Start studying machine learning

July 20

None

July 21

None

July 22

Pfu PCR & gel electrophoresis

  • Sample:ydfD, hipA,LuxI-LuxR, GFP-Ampicillin, mCherry.

Find dataset of mutation

July 23

Pfu PCR & gel electrophoresis

  • Sample: LuxI-LuxR, GFP-Ampicillin, ccdB, yafQ, chpBK, hipA, LuxI-LuxR(without Pcon)

Tramsformation

  • Sample:pSB1C3, pSB1K3, pSB3C5 Backbone.

July 24

Pfu PCR & gel electrophoresis

  • Sample: LuxI-LuxR, mCherry, MazF, LuxI-LuxR(without Pcon)

Gel extraction

  • Sample:MazF, LuxI-LuxR, GFP-Ampicillin, mCherry

Study the mechanism of mutation

July 25

Pfu PCR & gel electrophoresis

  • Sample:ccdB, yafQ, ydfD, hipA, LuxI-LuxR(without Pcon)

    • Digestion

    • Sample:LuxI-LuxR, mCherry, MazF, pSB1A3

    Ligation

    • Sample:
      • LuxI-LuxR+pSB1A3
      • mCherry+pSB1A3
      • MazF+pSB1A3

    Transformation

    • Sample:
      • plasmid with LuxI-LuxR into DH5α
      • plasmid with mCherry into DH5α
      • plasmid with MazF into DH5α

    Gel extraction

    • Sample:ccdB, yafQ, ydfD, hipA

    July 26

    Pfu PCR & gel electrophoresis

  • Sample:LuxI-LuxR(without Pcon)

    • Digestion

    • Sample:LuxI-LuxR, mCherry, MazF, pSB1A3

    Ligation

    • Sample:
      • LuxI-LuxR+pSB1A3
      • mCherry+pSB1A3
      • MazF+pSB1A3

    July 27

    Transformation

    • Sample:
      • plasmid with ccdB into DH5α
      • plasmid with yafQ into DH5α
      • plasmid with ydfD into DH5α
      • plasmid with hipA into DH5α

    July 28

    Pfu PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon)

    July 29

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, yafQ, ydfD, hipA

    Gel extraction

    • Sample:ccdB, yafQ, ydfD, hipA

    Digestion

    • Sample:ccdB, yafQ, ydfD, hipA, pSB1C3

    Ligation

    • Sample:
      • ccdB+pSB1C3
      • yafQ+pSB1C3
      • ydfD+pSB1C3
      • hipA+pSB1C3

    July 30

    Transformation

    • Sample:
      • plasmid with ccdB into DH5α
      • plasmid with yafQ into DH5α
      • plasmid with ydfD into DH5α
      • plasmid with hipA into DH5α

    Pfu PCR & gel electrophoresis

    • Sample:chpBK

    Colony PCR & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, MazF transformation product

    July 31

    Pfu PCR & gel electrophoresis

    • Sample:chpBK

    Digestion

    • Sample:ccdB, yafQ, ydfD, hipA, pSB1A3

    Ligation

    • Sample:
      • ccdB+pSB1A3
      • yafQ+pSB1A3
      • ydfD+pSB1A3
      • hipA+pSB1A3

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, ccdB, ydfD transformation product

    August 1

    Transformation

    • Sample:
      • plasmid with ccdB into DH5α
      • plasmid with yafQ into DH5α
      • plasmid with ydfD into DH5α
      • plasmid with hipA into DH5α

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, ydfD

    Start training AI

    August 2

    Pfu PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon)

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon) (With new primer.)

    Culturing

    August 3

    Plasmid extraction & Sequencing

    August 4

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon)

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, yafQ

    Gel extraction

    • Sample:ccdB, yafQ gel electrophoesis product

    August 5

    Plasmid extraction & Sequencing

    • Sample:LuxI-LuxR, mCherry

    August 6

    Digestion

    • Sample:ccdB, yafQ

    August 7

    Digestion

    • Sample:pSB1A3

    Ligation

    • Sample:
      • ccdB+pSB1A3
      • yafQ+pSB1A3

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon)

    Transformation

    • Sample:
      • Plasmid with ccdB into Rosetta gami
      • Plasmid with yafQ into Rosetta gami

    August 8

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR(without Pcon)

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, yafQ

    Gel extraction

    • Sample:ccdB, yafQ, LuxI-LuxR(without Pcon) gel electrophoresis product

    Digestion

    • Sample:ccdB, yafQ, LuxI-LuxR(without Pcon), pSB1A3

    Improving accuracy

    August 9

    Ligation

    • Sample:LuxI-LuxR(without Pcon)
      • ccdB+pSB1A3
      • yafQ+pSB1A3
      • LuxI-LuxR(without Pcon)+pSB1A3

    Transformation

    • Sample:
      • Plasmid with ccdB into Rosetta gami
      • Plasmid with yafQ into Rosetta gami
      • Plasmid with LuxI-LuxR(without Pcon) into Rosetta gami

    Start using Keras

    August 10

    Digestion

    • Sample:ccdB, yafQ, 46, pSB1A3

    Ligation

    • Sample:
      • ccdB+pSB1A3
      • yafQ+pSB1A3
      • LuxI-LuxR(without Pcon)+pSB1A3

    Colony PCR & gel electrophoresis

    • Sample:
      • plasmid with ccdB.
      • plasmid with yafQ.

    August 11

    Transformation

    • Sample:
      • Plasmid with ccdB into Rosetta gami.
      • Plasmid with yafQ into Rosetta gami.
      • Plasmid with LuxI-LuxR(without Pcon) into DH5α.

    Pfu PCR & gel electrophoresis

    • Sample:MazF, ydfD, chpBK, hipA.

    Gel extraction

    • Sample:MazF, ydfD, chpBK, hipA gel electrophoresis product.

    August 12

    Digestion

    • Sample:MazF, ydfD, chpBK, pSB1A3.

    Ligation

    • Sample:
      • MazF+pSB1A3
      • ydfD+pSB1A3
      • chpBK+pSB1A3

      Pfu PCR & gel electrophoresis

      • Sample:
        • hipA+pSB1A3
        • LuxI-LuxR(without Pcon)+pSB1A3

      Gel extraction

      • Sample:LuxI-LuxR(without Pcon) gel electrophoresis product.

      Culturing

      Start studying Tensorflow

    August 13

    Digestion

    • Sample:MazF, ydfD, chpBK, pSB1A3.

    Culturing

    • Sample:
      • Rosetta gami with ccdB
      • Rosetta gami with yafQ

    Ligation

    • Sample:
      • MazF+pSB1A3
      • ydfD+pSB1A3
      • chpBK+pSB1A3

    August 14

    Colony PCR

    • Sample:LuxI-LuxR+mCherry.

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, hipA.

    Gel extraction

    • Sample:ccdB, hipA.

    Digestion

    • Sample:ccdB, hipA, LuxI-LuxR+mCherry, pSB1A3, pSB1K3.

    Transformation

    • Sample:
      • Plasmid with MazF into BL21 DE3.
      • Plasmid with ydfD into BL21 DE3.
      • Plasmid with chpBK into BL21 DE3.

    August 15

    Ligation

    • Sample:
      • ccdB+pSB1A3
      • hipA+pSB1A3
      • LuxI-LuxR+mCherry+pSB1K3

    Transformation

    • Sample:
      • Plasmid with ccdB into BL21 DE3.
      • Plasmid with hipA into BL21 DE3.
      • Plasmid with LuxI-LuxR+mCherry into DH5α.

    Culturing

    • Sample:
      • Culure with pSB1C3.
      • Culure with pSB1K3.
      • LuxI-LuxR(without Pcon).

    Digestion

    • Sample:pSB1K3

    Fix growth model

    August 16

    Plasmid extraction

    • Sample:Plasmid with LuxI-LuxR(without Pcon).

    Digestion

    • Sample:LuxI-LuxR, mCherry, pSB1K3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1K3

    Culturing

    • Sample:
      • Culure with MazF.
      • Culure with ydfD.
      • Culure with chpBK.

    Colony PCR & gel electrophoresis

    • Sample:LuxI-LuxR+mCherry, ccdB, hipA.

    August 17

    Culturing

    Plasmid extraction

    • Sample:
      • Plasmid with MazF.
      • Plasmid with ydfD.
      • Plasmid with chpBK.

    Digestion

    • Sample:pSB1C3

    Ligation

    • Sample:pSB1C3 backcone

    August 18

    Transformation

    • Sample:
      • pSB1C3 backcone into BL21 DE3.
      • LuxI-LuxR+mCherry+pSB1K3 into BL21 DE3.

    August 19

    Colony PCR & gel electrophoresis

    • Sample:
      • Plasmid with ccdB transformation product.
      • Plasmid with hipA transformation product.
      • Plasmid with LuxI-LuxR+mCherry transformation product.
      • Plasmid with pSB1C3 transformation product.

    August 20

    Taq PCR & gel electrophoresis

    • Sample:LuxI-LuxR+mCherry, ccdB, hipA, pSB1C3

    Digestion

    • Sample:LuxI-LuxR, mCherry, pSB1K3

    August 21

    Ligation

    • Sample:LuxI-LuxR+mCherry with pSB1K3

    Digestion & gel electrophoresis

    • Sample:pSB1C3 gel electrophoresis product.

    Gel extraction

    • Sample:pSB1C3

    Ligation

    • Sample:pSB1C3 backbon

    Transformation

    • Sample:LuxI-LuxR+mCherry+pSB1K3 into DH5α.

    August 22

    Taq PCR & gel electrophoresis

    • Sample:
      • ccdB.
      • hipA.
      • LuxI-LuxR+mCherry.
      • pSB1C3.
      • Beta carotene+pSB1A3.

    Pfu PCR & gel electrophoresis

    • Sample:
      • ccdB.
      • hipA.

    Ligation

    • Sample:LuxI-LuxR+mCherry with pSB1K3

    Gel extraction

    • Sample:ccdB, hipA.

    Culturing

    • Sample:hipA, K274000, pSB1C3.

    Feature extraction

    August 23

    Plasmid extraction

    • Sample:
      • Plasmid with part hipA.
      • Plasmid with part K274000.
      • Plasmid with part pSB1C3.

    Digestion

    • Sample:LuxI-LuxR, mCherry, ccdB, hipA, pSB1A3, pSB1K3.

    Ligation

    • Sample:
      • ccdB+pSB1A3.
      • hipA+pSB1A3.

    Transformation

    • Sample:
      • ccdB into BL21.
      • hipA into BL21.

    Gel electrophoresis & extraction

    • Sample:LuxI-LuxR, mCherry.

    Ligation

    • Sample:LuxI-LuxR+mCherry with pSB1K3

    August 24

    Transformation

    • Sample:LuxI-LuxR+mCherry with pSB1K3 into DH5α.

    August 25

    Digestion

    • Sample:LuxI-LuxR, mCherry, pSB1C3.

    Transformation

    • Sample:
      • K592024 into BL21.
      • K1033917 into BL21.
      • K1633915 into BL21.
      • K864102 into BL21.

    Culturing

    • Sample:ccdB hipA.

    August 26

    Gel electrophoresis & extraction

    • Sample:LuxI-LuxR+mCherry.

    Ligation

    • Sample:LuxI-LuxR+mCherry with pSB1C3.

    Plasmid extraction

    • Sample:
      • Plasmid with part ccdB.
      • Plasmid with part hipA.

    Transformation

    • Sample:pSB1C3 with LuxI-LuxR+mCherry into DH5α.

    Culturing

    • Sample:Culture with part K274200.

    Fix QS model

    August 27

    Transformation

    • Sample:
      • ccdB into BL21.
      • MazF into BL21.
      • yafQ into BL21.
      • ydfD into BL21.
      • chpBK into BL21.
      • hipA into BL21.

    August 27

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    August 29

    Tramsformation

    • Sample:Plasmid with LuxI-LuxR+mCherry into BL21.

    Start using SVM & ANN

    August 30

    Colony PCR & gel electrophoresis

    • Sample:Plasmid with LuxI-LuxR+mCherry transformtion product

    August 31

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    September 1

    Toxin gene function test

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK, hipA.

    September 2

    Toxin gene function test

    • Sample:ccdB, MazF, YafQ, ydfD, chpBK.

    Tramsformation

    • Sample:Plasmid with LuxI-LuxR+mCherry into BL21.

    September 3

    Colony PCR & gel electrophoresis

    • Sample:Plasmid with LuxI-LuxR+mCherry transformtion product

    September 4

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    September 5

    Tramsformation

    • Sample:Plasmid with LuxI-LuxR+mCherry into BL21.

    Studying SVM scoring function

    September 6

    Colony PCR & gel electrophoresis

    • Sample:Plasmid with LuxI-LuxR+mCherry transformtion product

    September 7

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Gel extraction

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    Colony PCR & gel electrophoresis

    • Sample:
      • Plasmid with ccdB transformation product.
      • Plasmid with MazF transformation product.
      • Plasmid with yafQ transformation product.
      • Plasmid with ydfD transformation product.
      • Plasmid with chpBK transformation product.
      • Plasmid with hipA transformation product.
      • Plasmid with LuxI-LuxR+mCherry transformation product.

    September 8

    Transformation

    • Sample:Plasmid with LuxI-LuxR+mCherry into BL21.

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 9

    Digestion & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Ligation

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 10

    Transformation

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 11

    Colony PCR and electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 12

    Gel electrophoresis & gel extraction

    • Sample:
      • ccdB ligation product.
      • MazF ligation product.
      • yafQ ligation product.
      • ydfD ligation product.
      • hipA ligation product.

    Plasmid extraction

    • Sample:
      • ccdB transformation product.
      • MazF transformation product.
      • yafQ transformation product.
      • ydfD transformation product.
      • hipA transformation product.

    Toxin gene function test

    • Sample:ccdB, MazF, YafQ, ydfD, chpBK.

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Transformation

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Troubleshooting of FNR

    September 13

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 14

    Taq PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    September 15

    Toxin gene function test

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK, hipA.

    Taq PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Colony PCR & gel electrophoresis

    • Sample:ccdB, MazF, ydfD, hipA.

    September 16

    Quorum sensing function test

    SDS

    • Sample:pSB1C3, gfaspurple.

    September 17

    Digestion & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA, pSB1C3

    Ligation

    • Sample:
      • ccdB+pSB1C3.
      • MazF+pSB1C3.
      • yafQ+pSB1C3.
      • ydfD+pSB1C3.
      • chpBK+pSB1C3.
      • hipA+pSB1C3.

    Taq PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Pfu PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA.

    Transformation

    • Sample:Plasmid with LuxI-LuxR+mCherry into DH5α.

    Gfaspurple absorbance specturm

    September 18

    Transformation

    • Sample:
      • Gfaspurple into DH5a.
      • pSB1A3 backbone into DH5α.
      • pSB1C3 backbone into DH5α.
      • K274200+pSB1A3 into DH5α.

    September 19

    Toxin gene function test

    • Sample:ccdB, MazF, YafQ, ydfD, chpBK.

    Colony PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK, hipA.

    Taq PCR & gel electrophoresis

    • Sample:chpBK.

    September 20

    Digestion & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, hipA, pSB1C3

    Ligation

    • Sample:
      • ccdB+pSB1C3.
      • MazF+pSB1C3.
      • yafQ+pSB1C3.
      • ydfD+pSB1C3.
      • chpBK+pSB1C3.
      • hipA+pSB1C3.

    Data balancing

    September 21

    Plasmid extraction

    • Sample:ccdB, MazF, yafQ, ydfD, hipA

    Toxin gene function test

    • ccdB, MazF, YafQ, ydfD, chpBK.

    Colony PCR & gel electrophoresis

    • Sample:RFP, PFP

    Transformation

    • Sample:
      • Plasmid with ccdB into BL21.
      • Plasmid with MazF into BL21.
      • Plasmid with yafQ into BL21.
      • Plasmid with ydfD into BL21.
      • Plasmid with chpBK into BL21.
      • Plasmid with hipA into BL21.

    Make competent cell & testing.

    • Competent cell type:BL21.

    Design UI of AI

    September 22

    Quorum sensing function test.

    Colony PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK.

    Colony PCR & gel electrophoresis

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK.

    Import pubchempy

    September 23

    Gel electrophoresis.

    • Sample:k864102，k592024, gfaspurple.

    September 24

    Chemical test

    • Sample:MazF, ydfD, chpBK
    • Chemical:NaN3
    • Concentration:1μM

    September 25

    Cotransformation.

    • Sample:Plasmid with LuxI-LuxR+mCherry & plasmid with ydfD into BL21.
    • Sample:Plasmid with LuxI-LuxR+mCherry & Plasmid with hipA into BL21.

    Colony PCR.

    • Sample:hipA

    September 26

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    September 27

    Quorum sensing function test

    Transformation

    • Sample:plasmisd with LuxI-LuxR+mCherry into BL21.

    September 28

    Transformation

    • Sample:plasmisd with J0yafQ1 into DH5α.

    Cotransformation

    • Sample:
      • plasmisd with LuxI-LuxR+mCherry & plasmid with ydfD into BL21.
      • plasmisd with LuxI-LuxR+mCherry & plasmid with chpBK into BL21.

    Colony PCR & gel electrophoresis

    • Sample:plasmisd with LuxI-LuxR+mCherry transformation product.

    September 29

    Toxin gene function test

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK, hipA.

    September 30

    Digestion & gel electrophoresis

    • Sample:LuxI-LuxR, mCherry, pSB1C3

    Ligation

    • Sample:LuxI-LuxR+mCherry+pSB1C3

    October 1

    Transformation

    • Sample:
      • plasmisd with J04500 into DH5α.
      • plasmisd with LuxI-LuxR+mCherry into BL21.

    Chemical test

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK.
    • Chemical:EtBr
    • Concentration:0.1μM

    October 2

    Plasmid extraction

    • Sample:ccdB, MazF, yafQ, ydfD, chpBK.

    Colony PCR

    • Sample:
      • plasmisd with J04500 transformation product.
      • plasmisd with LuxI-LuxR+mCherry transformation product.

    October 3

    Toxin gene function test

    • Sample:ydfD, chpBK.

    Chemical test

    • Sample:ydfD.
    • Chemical:EtBr
    • Concentration:0.05μM, 0.5μM, 1μM.

    October 4

    Chemical test

    • Sample:ydfD, chpBK.
    • Chemical:EtBr
    • Concentration:0.05μM, 0.5μM, 1μM.

    October 5

    Chemical test

    • Sample:ydfD.
    • Chemical:EtBr
    • Concentration:0.05μM, 0.5μM, 1μM.

    UV test

    • Sample:ydfD.
    • UV type:UVB
    • Strength:1mW/cm2, 2mW/cm2, 3mW/cm2.

    Transformation

    • Sample:Plasmid with ydfD into BL21.

    Toxin gene function test

    • Sample:ccdB, MazF, YafQ, ydfD, chpBK.

    Colony PCR & gel electrophoresis

    • Sample:MazF, yafQ, ydfD, YFP.

    Make competent cell

    • Sample:QS system.

    October 6

    Plasmid extraction

    • Sample:ydfD.

    Transformation

    • Sample:
      • Plasmid with ccdB into BL21.
      • Plasmid with MazF into BL21.
      • Plasmid with yafQ into BL21.
      • Plasmid with ydfD into BL21.
      • Plasmid with chpBK into BL21.

    Cotransformation

    • Sample:Plasmid with LuxI-LuxR+mCherry & plasmid ith ydfD into BL21.

    Colony PCR & gel electrophoresis

    • Sample:
      • Plasmid with LuxI-LuxR+mCherry transformation product.
      • Plasmid with ydfD transformation product.

    Chemical test

    • Sample:ydfD.
    • Chemical:EtBr
    • Concentration:
      • 0.5uM, 1uM.
      • 0.05uM, 0.5uM, 1uM.
      • 0.05uM, 0.1uM, 0.5uM, 1uM.

    UV test

    • Sample:ydfD.
    • UV type:UVB
    • Strength:1mW/cm2, 2mW/cm2, 3mW/cm2.

    October 7

    Cotransformation

    • Sample:Plasmid with LuxI-LuxR+mCherry & plasmid ith ydfD into BL21.

    Colony PCR & gel electrophoresis

    • Sample:QS-ydfD cotransformation product.

    QS system function test

    October 8

    Colony PCR & gel electrophoresis

    • Sample:QS, ccdB, MazF, yafQ, ydfD, chpBK.

    October 9

    Cotransformation function test

    • Sample:QS-ydfD.

    Toxin gene function test

    • Sample:ccdB, MazF, YafQ, ydfD, chpBK.

    October 10

    Cotransformation function test

    • Sample:QS-ydfD.

    October 11

    Transformation

    • Sample:BBa_I6060 into BL21.

    October 12

    UV & EtBr mutagenicity test

    • Sample:BL21 with ydfD inserted.

    October 13

    Toxin gene function test

    • Sample:ccdB, YafQ, ydfD, chpBK.

    October 14

    UV & EtBr mutagenicity test

    • Sample:ccdB, YafQ, ydfD, chpBK.

    October 15

    Colony PCR

    • Sample:BL21 with BBa_I6060 inserted.

    SDS Page

    • Sample:BL21 with BBa_I6060 inserted.

    Characterization Experiment

    • Sample:BBa_I6060.

    October 16

    UV & EtBr mutagenicity test

    • Sample:ccdB, YafQ, ydfD, chpBK.

    October 17

    Transformation

    • Sample:ccdB into BL21.

    October 18

    Toxin gene function test

    • Sample:ccdB.

    October 19

    EtBr mutagenicity test

    • Sample:ccdB.