Team:NCTU Formosa/Notebook

From 2014.igem.org

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Revision as of 12:39, 9 October 2014

Notebook

Contents

About the notes

Our notes are the abbreviations of the experimental process, including titles, genes we transformed, plasmids we used, and resistances on the plasmids. We categorized these notes by date. Moreover, team members followed the iGEM protocol in each experimental step in order to get better experimental efficiency and the results we had expected.

Material and Methods

Wet lab protocol

Insect experiment protocol

April 2014

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None

4/11

Transformation of [PSB1K3] Kr, [PSB1A3] Ar, [PSB1C3] Cr (G2)

Transformation of [Plac+PSB1A3] Ar, [mGFP+PSB1A3] Ar (G3)

Transformation of [PSB1K3] Kr,[PSB1A3] Ar, [PSB1C3] Cr (G1)

4/12

Cultivation of [PSB1K3] Kr, [PSB1C3] Cr (G2)

Cultivation of [PSB1A3] Ar, [PSB1C3] Cr (G1)

Cultivation of [mGFP] Ar (G3)


4/13

Mini-prep of [mGFP] Ar, [PSB1K3] Kr (G3)

Mini-prep of [PSB1K3] Kr, [PSB1C3] Cr (G2)

Mini-prep of [PSB1A3] Ar, [PSB1C3] Cr (G1)

4/17

Transformation of [PSB1K3] Kr (G1)

4/18

Cultivation of [PSB1K3] Kr (G1)

4/19

Transformation of [Plac+PSB1A3] Ar, [PSB1K3] Kr, [PSB1A3] Ar, [PSB1C3] Cr (G3)

Mini-prep of [PSB1K3] Kr (G1)


4/20

Transformation of [PSB1A3] Ar (G2)

Electrophoresis of [PSB1K3] Kr, [PSB1C3] Cr plasmid (G2)

4/21

Cultivation of [PSB1K3] Kr, [PSB1A3] Ar, [PSB1C3] Cr (G2)

4/22

Mini-prep of [PSB1K3] Kr, [PSB1A3] Ar, [PSB1C3] Cr (G2)


4/28

Electrophoresis of digested [PSB1K3] Kr(EP), [PSB1C3] Cr(EP),[PSB1A3] Ar(EP) (G1)

Cultivation of [PSB1C3] Cr (G1)

4/29

Mini-prep of [PSB1C3] Cr (G1)

Mini-prep of [PSB1C3] Cr (G2)

Transformation of [B0032] Ar, and [BFP] Cr (G2)

Cultivation of [PSB1K3] Kr, [PSB1A3] Ar (G1)

Cultivation of [B0032] Ar, and [BFP] Cr (G2)

4/30

Mini-prep of [PSB1A3] Ar (G1)

Mini-prep of [B0032] Ar, and [BFP] Cr (G2)

Transformation of [B0032] Ar, and [BFP] Cr, [T7promoter] Ar Kr and [T7 polymerase] Cr (G2)

Transformation of [J23101] Ar,[K592100] Cr,[B0032] Ar,[B0015] Ar (G1)

Digestion of [PSBIC3] Cr(EP), [B0032] Ar(ES), [BFP] Cr(XP) mini (G2)

Electrophoresis of plasmid and digested prouct of[PSBIC3] Cr(EP), [B0032] Ar(ES), [BFP] Cr(XP) mini (G2)


May 2014

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5/1

Cultivation of [PSB1C3]Cr, [BFP]Cr, [T7promoter]ArKr, [T7 polymerase]Cr and [B0015]Ar (G2)

Transformation of [T7 polymerase]Cr (G2)

5/2

Cultivation of [J23101] Ar,[K592100] Cr,[B0032] Ar,[B0015] Ar (G1)

Mini-prep of [PSB1C3]Cr, [BFP]Cr, [T7promoter]ArKr, [T7 polymerase]Cr and [B0015]Ar (G2)

Digestion of [PSB1C3]Cr(EP), [BFP]Cr(XP), [T7 polymerase]Cr(ES), [T7promoter]ArKr(ES) and [B0015]Ar*2(ES & XP) (G2)

5/3

Mini-prep of [J23101] Ar,[K592100] Cr,[B0032] Ar,[B0015] Ar (G1)

Mini-pep of [B0032] Ar, [B0015] Ar,[BFP] Cr (G3)

Digestion of [J23101] Ar (ES),[K592100]Cr(ES),[B0032] Ar(XP),[B0015] Ar(XP),[PSB1A3] Ar(EP),[PSB1C3] Cr(EP),[PSB1K3] Kr(EP) mini (G1)

Electrophoresis of plasmid and digested product of [PSB1C3]Cr(EP), [BFP]Cr(XP), [T7 polymerase]Cr(ES), [T7promoter]ArKr(ES) and B0015*2(ES & XP) (G2)

Electrophoresis of digested product of [J23101] Ar (ES),[K592100] Cr(ES),[B0032] Ar(XP),[B0015] Ar(XP),[PSB1A3] Ar(EP),[PSB1C3]Cr(EP),[PSB1K3] Kr(EP) (G1)

Cultivation of [B0015] Ar, [PSB1A3] Ar, [PSB1K3] Kr (G1)

Ligation of [BFP+B0032+PSB1K3]Kr (G2)

Ligation of [J23101+B0032+PSB1C3] Cr (G1)

Transformation of ligation product [BFP+B0032+PSB1K3]Kr (G2)

Transformation of [T7polymerase]Cr and [B0015]Ar (G2)

5/4

Mini-prep of [B0015] Ar, [PSB1A3] Ar, [PSB1K3] Kr (G1)

Mini-prep of [B0015*3] Ar and [T7polymerase] Cr (G2)

Digestion of [B0015] Ar(XP), [PSB1A3] Ar(EP), [PSB1K3] Kr (EP)(G1)

Digestion of [B0015*3] Ar and [T7polymerase] Cr (G2)

Digestion of [B0015] Ar(XP), [BFP] Cr(XP), [PSB1A3] Ar(EP), [PSB1C3] Cr(EP), [B0032] Ar (XP) (G3)

Transformation of [Pcons promoter] Ar, [RBS+BFP] Kr and [T7 promoter] Kr (G2)

Transformation of [LacI] Ar (G3)

Electrophoresis of plasmid and digested product of [B0015*3] Ar and [T7polymerase]Cr (G2)

Electrophoresis of digested product of [B0015] Ar(XP), [PSB1A3] Ar(EP), [PSB1K3] Kr (EP), [K592100] Cr (ES)(G1)

Cultivation of [B0015] Ar, [PSB1A3] Ar(G1)

Cultivation of [Plac] Ar, [PSB1C3] Cr, [mGFP] Ar (G3)

5/5

Mini-prep of [PSB1A3] Ar,[B0015]Ar (G1)

Mini-prep of [Plac] Ar, [PSB1C3] Cr, [mGFP] Ar (G3)

Digestion of [PSB1A3] Ar(EP),[B0015]Ar(XP)(G1)

Electrophoresis of digested product of [PSB1A3] Ar(EP),[B0015]Ar(XP),[PSB1K3]Kr(EP) (G1)

5/6

Mini-prep of [LacI] Ar (G3)

5/7

Transformation of [PT7polymerase]Cr and [J61048]Ar (G2)

5/8

Transformation of [B0015*3] Ar and [J61048*2] Ar (G2)

Transformation of [B0015] Ar and [J61048] Ar (G3)

Transformation of [B0015]Ar,[J61048]Ar,[J23101]Ar (G1)

Mini-prep of [PT7polymerase]Cr, [T7polymerase*2]Cr and [B0015*2]Ar (G2)

Digestion of [T7polymerase] Kr (EP) and [B0015] Ar (ES) (G2)

Digestion of [Plac] Ar (ES), [mGFP] Ar (ES), [LacI] Ar (ES), [PSB1C3] Cr (EP) (G3)

Electrophoresis of digested product of [Plac] Ar (ES), [mGFP] Ar (ES), [LacI] Ar (ES), [PSB1C3] Cr (EP) (G3)

5/9

Cultivation of [B0015] Ar, [J61048] Ar [LacI] Ar (G3)

5/10

Mini-prep of [LacI] Ar (G3)


5/11

Cultivation of [J61048] Ar (G2) (ES)

Electrophoresis of plasmid and digested product of [B0015]Ar(ES), [T7polymerase] Kr(only mini) and [T7promoter] Kr (EP)

Transformation of [T7 promoter] Kr (G2)

5/12

Digestion of [T7polymerase] Kr (ES) and [J61048] Ar (ES) (G2)

5/13

Transformation of [T7 promoter] Kr (G2)

5/15

Electrophoresis of plasmid and digested product of [J61048]Ar (ES), [T7polymerase*3]Kr(ES & XP) and [PSBIK3]Kr(EP) (G2)

Cultivation of [B0015]Ar, [J61048]Ar, [J23101]Ar, [PSB1K3]Kr (G1)

Mini-prep of [PSB1K3]Kr (G1)

Digestion of [PSB1K3]Kr (EP) (G1)


5/19

Cultivation of [J23101]Ar,[B0015]Ar,[J61048]Ar (G1)

5/20

Transformation of [T7polymerase] Kr and [PSB1K3] Kr (G2)

Electrophoresis of plasmid and digested product of [T7polymerase*6] Kr and [PSB1K3*4]Kr (G2)

Mini-prep of [J23101]Ar, [B0015]Ar, [J61048]Ar (G1)

Digestion of [J23101]Ar (ES), [B0015]Ar (XP), [J61048]Ar (XP) (G1)

5/21

Digestion of [T7polymerase*6] Kr and [PSB1K3*2] Kr (G2)

Mini-prep of [T7polymerase] Kr and [PSB1K3] Kr (G2)

Electrophoresis of plasmid and digested product of [J23101]Ar(ES), [B0015]Ar(XP), [J61048]Ar(XP) and digested product [PSB1K3]Kr(EP) (G1)

Ligation of [K592100+J61048+PSB1K3] Kr (G1)

5/22

Electrophoresis of plasmid and digested product of [T7polymerase] Kr (ES) and [PSB1K3]Kr (EP) (G2)

5/23

Transformation of ligation product [K592100+J61048+PSB1K3]Kr (G1)

5/24

Digestion of [J61048] Ar (XP) (G2)

PCR of [BFP+B0032*4] Kr and [T7 promoter*1] Ar Kr (G2)

Electrophoresis of PCR products [BFP+B0032*4] Kr and [T7 promoter*1] Ar Kr, [J61048] Ar (G2)


5/27

Electrophoresis of digested product of [mGFP] Ar (ES), [J61048] Ar (XP) ,[B0015] Ar (XP) (G3)

Digestion of [J61048] Ar (XP) and [T7polymerase] Kr (ES) (G2)

Cultivation of [PSB1K3*3] Kr(G2)

5/28

PCR of [PSBIK3*5]Kr , [PSBIA3*5]Ar , [PSBIC3*5]Cr ,[J61048*5]Ar ,[B0015*5]Ar and [BFP*5](G1)

Mini-prep of [PSB1K3*3] Kr(G2)

Digestion of [PSB1K3*3] Kr (EP) (G2)

Electrophoresis of plasmid and digested product of [PSB1K3] Kr (EP), [T7polymerase] Kr (ES), [J61048] Ar (XP) (G2)

5/29

Digestion of [mGFP] Ar (ES), [J61048] Ar (XP) ,[B0015] Ar (XP) (G3)

Transformation of [J23101] Ar (G1)

Transformation of [T7 promoter] Ar Kr (G2)

Electrophoresis of plasmid and digested product of [J61048] Ar (ES), [T7polymerase] Kr (ES) (G2)

Ligation of [J61048+J61048] Cr, [T7polymerase+J61048] Kr, [RBS+BFP] Kr (G2)

Ligation of [Plac+B0032+PSB1K3] Kr (G3)

5/30

Transformation of ligation product (G2)

Transformation of ligation product [Plac+B0032+PSB1K3] Kr (G3)


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6/2

PCR of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2)

Cultivation of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2)

Electrophoresis of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3]Kr and [B0032+BFP*3] Kr plasmid (G2)

6/3

PCR of [T7 promoter*5] Ar Kr, [B0032+BFP*5] Kr , [J61048+J61048*5] Cr and [T7polymerase+J61048*5] Kr (G2)

Electrophoresis of PCR products (G2)

Cultivation of [B0032+BFP*10] Kr, [J61048+J61048*10] Cr and [T7polymerase+J61048*5] Kr (G2)

6/5

Transformation of ligation product [K592100+J61048+PSBIK3] Kr (G1)

Ligation of [BFP+B0032] Kr and [T7polymerase+J61048] Kr (G2)

Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)

6/6

Mini-prep of [J61048+J61048*3] Cr and [T7 promoter*3]Ar Kr (G2)

Digestion of [J61048+J61048*3] Cr (XP) and [T7 promoter*3]Ar Kr(ES) (G2)

Transformation of [B0032+BFP] Kr and [T7polymerase+J61048] Kr (G2)

Transformation of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)

6/7

PCR of [PSB1K3*6] Kr, [K592100+J61048+PSB1K3*10] Kr,[J23101*8] Ar ,[B0032*8] Ar (G1)

PCR of [mGFP+B0015+PSB1K3*6] Kr and [mGFP+J61048+PSB1K3*6] Kr (G3)


6/8

Electrophoresis of plasmid and digestion product of [J61048+J61048](XP), [T7 promoter](ES)

PCR of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)

Cultivation of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)

Electrophoresis of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)

6/9

Transformation of [B0023*2 ] Ar ,[J23101*2] Ar, [K592100+J61048*2] Kr and [PSB1K3*2] Kr (G1)

Electrophoresis of plasmid and digestion product of [RBS+BFP]Kr (ES),[T7 polymerase+J61048]Kr(XP),[T7 promoter]Kr Cr (ES),[J61048+J61048]Cr(XP) (G2)

Mini-prep of the plasmid and then digested the plasmid of [T7 polymerase+J61048]Kr(XP),[RBS+BFP]Kr(ES) (G2)

6/10

Electrophoresis of plasmid and digestion product of [K592100+J61048*2] Kr (XP), [PSB1K3*2] Kr (EP),[J23101*2] Ar (SP) and [B0023*2 ] Ar (SP) (G1)

6/11

Digestion of [mGFP] Ar (ES), [J61048] Ar (SP) ,[B0015] Ar (SP) (G3)

6/12

Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)

PCR of [Plac+B0032+PSB1K3*12] Kr (G3)

6/13

Transformation of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)

Cultivation of [Plac+B0032+PSB1K3*4] Kr (G3)

6/14

Mini-prep of [Plac+B0032+PSB1K3*2] Kr (G3)

Cultivation of [Plac+B0032+PSB1K3*4] Kr (G3)

PCR of [mGFP+B0015+PSB1K3*6] Kr


6/15

Mini-prep of [Plac+B0032+PSB1K3*4] Kr (G3)

Digestion of [Plac+B0032+PSB1K3*4] Kr (ES) (G3)


6/24

Digestion of [J23101] Ar (ES) and [B0032] Ar (XP) (G1)

Ligation of [B0032+K592100+J61048] Ar (G1)

Electrophoresis of plasmid and digestion product of [K592100+J61048] Kr (XP), [J23101] Ar (SP) and [B0032] Ar (SP) (G1)

Digestion of [T7 polymerase +J61048]Kr(XP),[J61048+J61048]Cr(XP) (G2)

6/27

Digestion of [J61048+J61048] Cr (XP) (G2)

Electrophoresis of [RBS+BFP]kr(ES)(mini),[T7polymerase](ES)(mini),[T7 promoter](mini)

6/28

Transformation of mini [B0032+BFP+J61048] Cr (G2)


6/29

PCR of [B0032+BFP+J61048]Cr

Electrophoresis of the PCR products (G2)


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7/1

Ligation of [J23101+B0032+PSB1C3] Cr (G1)

Digestion of [B0032] Ar

Electrophoresis of plasmid and digestion products of[B0032+BFP+J61048]Cr (XP) (G2)

7/2

Transformation of ligation product [B0032+K592100+J61048] Ar and [J23101+B0032] Cr (G1)

Cultivation of [B0032+BFP+J61048] Cr in LB broth (G2)

7/3

Mini-prep of [B0032+BFP+J61048] and then digestion: (XP)

Electrophoresis of plasmid and digestion products of[B0032+BFP+J61048]Cr (XP) (G2)

7/4

Digestion of [T7 promoter](SP) (G2)

Cultivation of [J23101] Ar (G3)

7/5

PCR of [B0032+K592100+J61048*11] Ar (G1)

Electrophoresis of PCR products of(G1)

Ligation of [J23101+B0032+PSB1C3*3] Cr (G1)

Electrophoresis of plasmid and digestion products of[T7promoter](SP),[B0031+BFP+J61048](XP)

Ligation of [T7 promoter])(SP)+[B0032+BFP+J61048](XP) (G2)


7/6

Transformation of the ligation products made at 7/5 (G2)

7/7

Transformation of [J23101+B0032*5] Cr (G1)

Cultivation of [B0032+K592100+J61048*2] Ar (G1)

Cultivation of [J23101] Ar (G3)

Digestion of [B0015] Ar (XP), [J61048] Ar (XP) (G3)

7/8

Mini-prep of [B0032+K592100+J61048*2] Ar and then digest of only one of two: (XP) (G1)

PCR of [T7+B0032+BFP+J61048] (G2)

Digestion of [J23101] Ar (ES) (G3)

7/9

Cultivation of [B0032+BFP+J61048]Cr and [T7+B0032+BFP+J61048]Kr(G2)

7/10

Mini-prep of [B0032+BFP+J61048]Cr

Digestion of [B0032+FP+J61048]Cr( XP)

Electrophoresis of plasmid and digestion products of[B0032+BFP+J61048]Cr (XP) (G2)

7/12

Digestion of [B0032+BFP+J612048](XP), [T7+B0032+BFP+J61048]Kr (XP) (G2)


7/14

Electrophoresis of [B0032+BFP+J61048]Cr,[T7+B0032+BFP+J61048]Kr (G2)

7/15

Cultivation of [B0032+BFP+J61048]Cr,[PT7+B0032+BFP+J61048]Kr E.coli in LB broth (G2)

7/16

Mini-prep of [B0032+BFP+J61048],[PT7+B0032+BFP+J61048]

Digestion of [B0032+BFP+J61048]Cr(XP),[PT7+B032+BFP+J61048]Kr(ES) (G2)

7/17

Electrophoresis of the plasmid and digestion products made yesterday (G2)

7/19

PCR of [J23101+B0032*12] Cr (G1)

Electrophoresis of PCR products of [B0032+K592100+J61048*2] Ar ,digestion product of [B0032+K592100+J61048] Ar (XP) and [J23101] Ar (SP) (G1)


7/20

Digestion of [B0032+K592100+J61048] Ar (EP) (G1)

Digestion of [J23101] Ar (ES) (G3)

Ligation of [B0032+K592100+J61048] Cr (G1)

Ligation of [J23101+B0032+PSB1K3] Kr (G3)

PCR of [J23101+B0032*4] Cr (G1)

Cultivation of [J61048+B0032*4]Cr E.coli in LB broth (G1)

Electrophoresis of the PCR products and digestion products of [J23101] Ar (SP) and [B0032+K592100+J61048] Ar (EP) (G1)

7/21

Transformation of ligation product made yesterday (G1)

Transformation of ligation product of [J23101+B0032+PSB1K3] Kr (G3)

Ligation of [mGFP+J61048+PSB1K3] Kr,[mGFP+B0015+PSB1K3] Kr (G3)

7/23

Cultivation of [B0032+K592100+J61048] Cr E.coli in LB broth (G1)

Digestion of [J23101+B0032*3] Cr (EP) (G1)

Digestion of [B0032+K592100+J61048] Ar (EP) (G1)

Ligation of [J23101+B0032] Cr (G1)

Mini-prep of [B0032+K592100+J61048] Cr

Transformation of [B0032+K592100+J61048] Cr and [J23101+B0032] Ar (G1)

7/24

PCR of [B0032+K592100+J61048*12] Cr (G1)

Electrophoresis of digestion products of[J23101+B0032*3] Cr (EP) and the PCR products (G1)

Transformation of [J23101+B0032] Cr (G1)

Digestion of [BFP] Cr (ES), [B0015] Ar (XP), [J61048] Ar (XP), [B0032] Ar (XP), [BFP+B00315*2] Kr (XP) (G3)

7/26

Cultivation of [J23101+B0032*4] Cr E.coli in LB broth (G1)

Digestion of [J23101] Ar (SP), [B0032] Ar (XP) (G3)


7/27

Ligation of [J23101+B0032+PSB1C3] Cr (G3)

7/28

Digestion of [B0032+K592100+J61048*2] Cr (XP) (G1)

Transformation of [J23101] Ar (G3)

PCR of [J23101+B0032+PSB1C3*6] Cr (G3)

7/29

Electrophoresis of the digestion products of[B0032+K592100+J61048*2] Cr (XP) , [PSB1K3*2] Kr (EP) , [PSB1A3*3] Ar (EP) , [PSB1C3] Cr (EP) , [J23101] Ar (ES) , [B0032] Ar (SP) , [B0032] Ar (XP) , [B0032+K592100+J61048] Ar (XP) and [J23101] Ar (SP) (G1)

Electrophoresis of the digestion product of [B0032] Ar (EP),[PSB1C3] Cr (EP), [PSB1K3] Kr (EP), [PSB1A3] Ar (EP) and PCR product [J23101*6] Ar (G3)

Digestion of [B0032+K592100+J61048*2] Cr (XP) (G1)

Digestion of [B0032] Ar (EP), [PSB1C3] Cr (EP), [PSB1K3] Kr (EP), [PSB1A3] Ar (EP) (G3)

Mini-prep of [PT7+B0032+BFP+J61048] (G2)

PCR of [J23101*6] Ar (G3)

7/30

Electrophoresis of the digestion products of [B0032+K592100+J61048*2] Cr (XP) , [PSB1K3*2] Kr (EP) , [PSB1A3*3] Ar (EP) , [PSB1C3] Cr (EP) , [J23101] Ar (ES) , [B0032] Ar (SP) , [B0032] Ar (XP) , [B0032+K592100+J61048] Ar (XP) and [J23101] Ar (SP) (G1)

Electrophoresis of the plasmid and digestion products of [RBS],[B0032+BFP+J61048](XP),[PT7+B0032+BFP+J61048](XP) (G2)

7/31

Digestion of [B0032+K592100+J61048*2] Cr (XP) and [J23101] Ar (EP) (G1)

Ligation of [J23101+B0032+PSB1C3] Cr (G1)

Digestion of [B0032+K592100+J61048*2] Cr (XP) (G1)

Electrophoresis of the plasmid and digestion products of [B0032+K592100+J61048*2] Cr (XP) ,digestion products made in 7/7 and 7/30[B0032+K592100+J61048*2] Cr (XP) ; digestion product made in 7/31[B0032+K592100+J61048*2] Ar (XP) (G1)


August 2014

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None

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8/11

Transformation of NCTU1 (G1)

Transformation of PBAN (G2)

8/12

Cultivation of [PBAN-4]Kr,[PBAN-5]Krand [PBAN-6]Kr E.coli in LB broth (G1)

Cultivation of [RBS+BFP+J61048]Cr ,[T7+RBS+BFP+J61048]Ar Kr

Mini-prep of [Pcons]Ar

Digestion of [Pcons]Ar (ES) (G2)

8/13

Mini-prep of [PBAN-4] Kr,[PBAN-5] Krand [PBAN-6] Kr (G1)

Digestion of [PBAN-4]Kr(EP),[PBAN-5]Kr(EP)and [PBAN-6]Kr(EP) (G1)

Electrophoresis of [Pcons](mini)&(dig ES),digestion of [Pcons](XP)

Ligation of [Pcons]+[RBS+BFP+J61048] (G2)

8/14

Electrophoresis of plasmid and digestion products of [PBAN-4] Kr(EP),[PBAN-5] Kr(EP)and [PBAN-6] Kr(EP) (G1)

Purification from electrophoresis of [PBAN]No.4&5&6(G1)

Transformation of the ligation product made yesterday

Electrophoresis of plasmid and digestion products of [PBAN]No.4&5&6,[Pcons],[Pcons+RBS+BFP+J61048] (G2)

8/15

Digestion of purification products yesterday:[PBAN]No.4&5&6 (EP) (G1)

Purification from electrophoresis of [PBAN]No.4&5&6

Transformation of [B0034]Ar (G2)

8/16

Electrophoresis of the purification products from electrophoresis,digestion products of[PBAN]No.4&5&6 (EP) (G1)

PCR of [B0034]

Electrophoresis of [PSB1K3](mini)&(dig E),[Pcons](dig SP),PCR products of B0034

Mini-prep of [B0034]Ar E.coli

Ligation of [Pcons]+[B0034] +PSB1K3

Digestion of [BFP](ES)

Electrophoresis of plasmid and digestion products of [PSB1K3],[PSB1C3],5/12[J61048],[Pcons])


8/17

Electrophoresis of the digestion products of[PBAN]No.4&5&6 (EP) (G1)


None

None

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