Team:NCTU Formosa/Notebook/test2

5/1

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

Transform 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: [BFP+B0032+PSB1K3]Kr (G2)

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

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

Transform 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)

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

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

5/6 Mini-prep of [LacI] Ar (G3)

5/7 Transform of [PT7polymerase]Cr and [J61048]Ar (G2)

5/8 Transform of [B0015*3] Ar and [J61048*2] Ar (G2) Transform of [B0015] Ar and [J61048] Ar (G3) Transform 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 [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) Transform of [T7 promoter] Kr (G2)

5/12 Digestion of [T7polymerase] Kr (ES) and [J61048] Ar (ES) (G2)

5/13 Transform 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 Transform 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 digested product and plasmid [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 Transform of ligation product [K592100+J61048+PSB1K3]Kr (G1)

5/24 Digestion of [J61048] Ar (XP) (G2) PCR: [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:[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 [PSB1K3] Kr (EP), [T7polymerase] Kr (ES), [J61048] Ar (XP) (G2)

5/29 Digestion of [mGFP] Ar (ES), [J61048] Ar (XP) ,[B0015] Ar (XP) (G3) Transform of [J23101] Ar (G1) Transform 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 Transform of ligation product (G2) Transform of ligation product [Plac+B0032+PSB1K3] Kr (G3)