Team:NCTU Formosa/Notebook

From 2014.igem.org

(Difference between revisions)
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====Wet lab protocol====
====Wet lab protocol====
<ul class="download">  
<ul class="download">  
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<li>Protocol<p></p>English version download</li>
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<li>Protocol<p></p>[https://drive.google.com/file/d/0B3AZGZCnvCVCT0NaR1ozMnpieVk/view  English version download]</li>
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<li>實驗手冊<p></p>[https://drive.google.com/file/d/0B3AZGZCnvCVCT0NaR1ozMnpieVk/view  中文版本下載]</li>
+
<li>實驗手冊<p></p>[https://drive.google.com/file/d/0B3AZGZCnvCVCTndIYmloeENVVTA/view  中文版本下載]</li>

Revision as of 13:51, 2 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

  • Sun.
  • Mon.
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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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