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Team:NCTU Formosa/Notebook/test3
From 2014.igem.org
| Line 61: | Line 61: | ||
<p> | <p> | ||
| - | 6/2 | + | 6/2</p> |
| - | PCR: [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2) | + | <p> |
| - | Cultivation of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2) | + | PCR: [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2)</p> |
| - | Electrophoresis of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3]Kr and [B0032+BFP*3] Kr plasmid (G2) </p> | + | <p> |
| + | Cultivation of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3] Kr and [B0032+BFP*3] Kr (G2)</p> | ||
| + | <p> | ||
| + | Electrophoresis of [T7 promoter*4] Ar Kr, [J61048+J61048*3] Cr, [T7polymerase+J61048*3]Kr and [B0032+BFP*3] Kr plasmid (G2)</p> | ||
| + | <p> | ||
| + | 6/3</p> | ||
| + | <p> | ||
| + | PCR: [T7 promoter*5] Ar Kr, [B0032+BFP*5] Kr , [J61048+J61048*5] Cr and [T7polymerase+J61048*5] Kr (G2)</p> | ||
| + | <p> | ||
| + | Electrophoresis of PCR products (G2)</p> | ||
<p> | <p> | ||
| - | |||
| - | |||
| - | |||
Cultivation of [B0032+BFP*10] Kr, [J61048+J61048*10] Cr and [T7polymerase+J61048*5] Kr (G2)</p> | Cultivation of [B0032+BFP*10] Kr, [J61048+J61048*10] Cr and [T7polymerase+J61048*5] Kr (G2)</p> | ||
<p> | <p> | ||
| - | 6/5 | + | 6/5</p> |
| - | Transform of ligation product [K592100+J61048+PSBIK3] Kr (G1) | + | <p> |
| - | Ligation of [BFP+B0032] Kr and [T7polymerase+J61048] Kr (G2) | + | Transform of ligation product [K592100+J61048+PSBIK3] Kr (G1)</p> |
| + | <p> | ||
| + | Ligation of [BFP+B0032] Kr and [T7polymerase+J61048] Kr (G2)</p> | ||
| + | <p> | ||
Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | ||
<p> | <p> | ||
| - | 6/6 | + | 6/6</p> |
| - | Mini-prep of [J61048+J61048*3] Cr and [T7 promoter*3]Ar Kr (G2) | + | <p> |
| - | Digestion of [J61048+J61048*3] Cr (XP) and [T7 promoter*3]Ar Kr(ES) (G2) | + | Mini-prep of [J61048+J61048*3] Cr and [T7 promoter*3]Ar Kr (G2)</p> |
| - | Transform of [B0032+BFP] Kr and [T7polymerase+J61048] Kr (G2) | + | <p> |
| + | Digestion of [J61048+J61048*3] Cr (XP) and [T7 promoter*3]Ar Kr(ES) (G2)</p> | ||
| + | <p> | ||
| + | Transform of [B0032+BFP] Kr and [T7polymerase+J61048] Kr (G2)</p> | ||
| + | <p> | ||
Transform of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | Transform of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | ||
<p> | <p> | ||
| - | 6/7 | + | 6/7</p> |
| - | PCR: [PSB1K3*6] Kr, [K592100+J61048+PSB1K3*10] Kr,[J23101*8] Ar ,[B0032*8] Ar (G1) | + | <p> |
| + | PCR: [PSB1K3*6] Kr, [K592100+J61048+PSB1K3*10] Kr,[J23101*8] Ar ,[B0032*8] Ar (G1)</p> | ||
| + | <p> | ||
PCR: [mGFP+B0015+PSB1K3*6] Kr and [mGFP+J61048+PSB1K3*6] Kr (G3)</p> | PCR: [mGFP+B0015+PSB1K3*6] Kr and [mGFP+J61048+PSB1K3*6] Kr (G3)</p> | ||
| Line 89: | Line 104: | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 6/8 | + | 6/8</p> |
| - | Electrophoresis of plasmid and digestion product of [J61048+J61048](XP), [T7 promoter](ES) | + | <p> |
| - | PCR: [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2) | + | Electrophoresis of plasmid and digestion product of [J61048+J61048](XP), [T7 promoter](ES)</p> |
| - | Cultivation of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2) | + | <p> |
| + | PCR: [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)</p> | ||
| + | <p> | ||
| + | Cultivation of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)</p> | ||
| + | <p> | ||
Electrophoresis of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)</p> | Electrophoresis of [B0032+BFP*5] Kr and [T7polymerase+J61048*5] Kr (G2)</p> | ||
<p> | <p> | ||
| - | 6/9 | + | 6/9</p> |
| - | Transform of [B0023*2 ] Ar ,[J23101*2] Ar, [K592100+J61048*2] Kr and [PSB1K3*2] Kr (G1) | + | <p> |
| - | 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) | + | Transform of [B0023*2 ] Ar ,[J23101*2] Ar, [K592100+J61048*2] Kr and [PSB1K3*2] Kr (G1)</p> |
| + | <p> | ||
| + | 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)</p> | ||
| + | <p> | ||
Mini-prep of the plasmid and then digested the plasmid:[T7 polymerase+J61048]Kr(XP),[RBS+BFP]Kr(ES) (G2)</p> | Mini-prep of the plasmid and then digested the plasmid:[T7 polymerase+J61048]Kr(XP),[RBS+BFP]Kr(ES) (G2)</p> | ||
<p> | <p> | ||
| - | 6/10 | + | 6/10</p> |
| + | <p> | ||
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)</p> | 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)</p> | ||
<p> | <p> | ||
| - | 6/11 | + | 6/11</p> |
| + | <p> | ||
Digestion of [mGFP] Ar (ES), [J61048] Ar (SP) ,[B0015] Ar (SP) (G3)</p> | Digestion of [mGFP] Ar (ES), [J61048] Ar (SP) ,[B0015] Ar (SP) (G3)</p> | ||
<p> | <p> | ||
| - | 6/12 | + | 6/12</p> |
| - | Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3) | + | <p> |
| + | Ligation of [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | ||
| + | <p> | ||
PCR: [Plac+B0032+PSB1K3*12] Kr (G3)</p> | PCR: [Plac+B0032+PSB1K3*12] Kr (G3)</p> | ||
<p> | <p> | ||
| - | 6/13 | + | 6/13</p> |
| - | Transform of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3) | + | <p> |
| + | Transform of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)</p> | ||
| + | <p> | ||
| + | Cultivation of [Plac+B0032+PSB1K3*4] Kr (G3)</p> | ||
| + | <p> | ||
| + | 6/14</p> | ||
| + | <p> | ||
| + | Mini-prep of [Plac+B0032+PSB1K3*2] Kr (G3)</p> | ||
| + | <p> | ||
Cultivation of [Plac+B0032+PSB1K3*4] Kr (G3)</p> | Cultivation of [Plac+B0032+PSB1K3*4] Kr (G3)</p> | ||
<p> | <p> | ||
| - | |||
| - | |||
| - | |||
PCR: [mGFP+B0015+PSB1K3*6] Kr </p> | PCR: [mGFP+B0015+PSB1K3*6] Kr </p> | ||
<br></div> | <br></div> | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 6/15 | + | 6/15</p> |
| - | Mini-prep [Plac+B0032+PSB1K3*4] Kr (G3) | + | <p> |
| + | Mini-prep [Plac+B0032+PSB1K3*4] Kr (G3)</p> | ||
| + | <p> | ||
Digestion of [Plac+B0032+PSB1K3*4] Kr (ES) (G3)</p> | Digestion of [Plac+B0032+PSB1K3*4] Kr (ES) (G3)</p> | ||
<br></div> | <br></div> | ||
| Line 128: | Line 161: | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 6/24 | + | 6/24</p> |
| - | Digestion of [J23101] Ar (ES) and [B0032] Ar (XP) (G1) | + | <p> |
| - | Ligation of [B0032+K592100+J61048] Ar (G1) | + | Digestion of [J23101] Ar (ES) and [B0032] Ar (XP) (G1)</p> |
| - | Electrophoresis of plasmid and digestion product of [K592100+J61048] Kr (XP), [J23101] Ar (SP) and [B0032] Ar (SP) (G1) | + | <p> |
| + | Ligation of [B0032+K592100+J61048] Ar (G1)</p> | ||
| + | <p> | ||
| + | Electrophoresis of plasmid and digestion product of [K592100+J61048] Kr (XP), [J23101] Ar (SP) and [B0032] Ar (SP) (G1)</p> | ||
| + | <p> | ||
Digestion of [T7 polymerase +J61048]Kr(XP),[J61048+J61048]Cr(XP) (G2)</p> | Digestion of [T7 polymerase +J61048]Kr(XP),[J61048+J61048]Cr(XP) (G2)</p> | ||
<p> | <p> | ||
| - | 6/27 | + | 6/27</p> |
| - | Digestion of [J61048+J61048] Cr (XP) (G2) | + | <p> |
| + | Digestion of [J61048+J61048] Cr (XP) (G2)</p> | ||
| + | <p> | ||
Electrophoresis of [RBS+BFP]kr(ES)(mini).[T7polymerase](ES)(mini).[T7 promoter](mini)</p> | Electrophoresis of [RBS+BFP]kr(ES)(mini).[T7polymerase](ES)(mini).[T7 promoter](mini)</p> | ||
<p> | <p> | ||
| - | 6/28 | + | 6/28</p> |
| + | <p> | ||
Transformation of mini:[B0032+BFP+J61048] Cr (G2)</p> | Transformation of mini:[B0032+BFP+J61048] Cr (G2)</p> | ||
<br></div> | <br></div> | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 6/29 | + | 6/29</p> |
| - | PCR:[B0032+BFP+J61048]Cr | + | <p> |
| + | PCR:[B0032+BFP+J61048]Cr</p> | ||
| + | <p> | ||
Electrophoresis of the PCR products (G2)</p> | Electrophoresis of the PCR products (G2)</p> | ||
<br></div> | <br></div> | ||
Revision as of 13:48, 13 October 2014
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6/2
PCR: [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: [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
Transform 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)
Transform of [B0032+BFP] Kr and [T7polymerase+J61048] Kr (G2)
Transform of ligation product [mGFP+B0015+PSB1K3] Kr and [mGFP+J61048+PSB1K3] Kr (G3)
6/7
PCR: [PSB1K3*6] Kr, [K592100+J61048+PSB1K3*10] Kr,[J23101*8] Ar ,[B0032*8] Ar (G1)
PCR: [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: [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
Transform 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:[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: [Plac+B0032+PSB1K3*12] Kr (G3)
6/13
Transform 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: [mGFP+B0015+PSB1K3*6] Kr
6/15
Mini-prep [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:[B0032+BFP+J61048]Cr
Electrophoresis of the PCR products (G2)
