Team:NCTU Formosa/Notebook/test7

From 2014.igem.org

(Difference between revisions)
Line 83: Line 83:
  [SacI-1]Cr</p><p>  
  [SacI-1]Cr</p><p>  
[AseI-1]Cr</p><p>
[AseI-1]Cr</p><p>
-
[ P<sub>cons</sub>+RBS+HindIII-1]Cr</p><p>
+
[P<sub>cons</sub>+RBS+HindIII-1]Cr</p><p>
-
  [ P<sub>cons</sub>+RBS+KpnI-4]Cr</p><p>
+
  [P<sub>cons</sub>+RBS+KpnI-4]Cr</p><p>
-
  [ P<sub>cons</sub>+RBS+AseI-5]Cr</p><p>  
+
  [P<sub>cons</sub>+RBS+AseI-5]Cr</p><p>  
-
[ P<sub>cons</sub>+RBS+XhoI-1]Cr</p><p>
+
[P<sub>cons</sub>+RBS+XhoI-1]Cr</p><p>
-
  [ P<sub>cons</sub>+RBS+SacI-1]Cr</p><p>  
+
  [P<sub>cons</sub>+RBS+SacI-1]Cr</p><p>  
-
[ P<sub>cons</sub>+RBS+KpnI-1+J61048]Cr</p><p>
+
[P<sub>cons</sub>+RBS+KpnI-1+J61048]Cr</p><p>
  [SalI-4]Cr</p><p>  
  [SalI-4]Cr</p><p>  
[P<sub>cons</sub>+RBS+SalI-3+J61048]Cr</p><p>
[P<sub>cons</sub>+RBS+SalI-3+J61048]Cr</p><p>

Revision as of 15:05, 17 October 2014

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10/3

Cultivation of

[Pcons+RBS+NdeI]Cr

[Pcons+RBS+NcoI]Cr

[KpnI-1]Cr

[NdeI-1]Cr

[XhoI-1]Cr

[SacI-1]Cr

[AseI-1]Cr

[Pcons+RBS+HindIII-1]Cr

[Pcons+RBS+KpnI-4]Cr

[Pcons+RBS+AseI-5]Cr

[Pcons+RBS+XhoI-1]Cr

[Pcons+RBS+SacI-1]Cr

[Pcons+RBS+KpnI-1+J61048]Cr

[SalI-4]Cr

[Pcons+RBS+SalI-3+J61048]Cr

[HindIII-4]Cr

[Pcons+RBS+AseI+J61048]Cr

[NheI]

[NcoI]

Electrophoresis of

[NdeI](EP)

[NcoI](EP)

[SacI](EP)

Transformation of

[Pcons+RBS+NdeI]Cr

[Pcons+RBS+SacI]Cr

DNA Sequencing of

[Pcons+RBS+NdeI+J61048-1]

[Pcons+RBS+HindIII+J61048-1]

[Pcons+RBS+SalI+J61048-4]

[Pcons+RBS+SacI+J61048-1]

[Pcons+RBS+SacI+J61048-1](fail)

10/4

Digestion of

[Pcons+RBS+XhoI+J61048-2, -4, -5]

[Pcons+RBS+SacI+J61048-10, -11]

[Pcons+RBS+NheI+J61048-20]

[Pcons+RBS+NheI]Cr

[Pcons+RBS+NcoI]Cr

Cultivation of

[Pcons+RBS+NheI-1, -2, -4, -6, -10, -12]

[Pcons+RBS+SalI-1, -3, -6, -7, -11, -12]

Cultivation of

[Pcons+RBS+NcoI+J61048-1~8]

PCR of

[Pcons+RBS+NheI-1, -2, -4, -6, -10, -12]

[Pcons+RBS+SacI-1, -3, -6, -7, -11, -12]

[Pcons+RBS+NcoI+J61048-1~8]

Ligation of

[Pcons+RBS+NheI]Kr

[Pcons+RBS+NcoI]Kr

Electrophoresis of

[XhoI-2, -4, -5](dig)

[SacI-10, -11](dig)

[NheI-20](dig)

[Pcons+RBS+NdeI](dig)

[Pcons+RBS+NcoI](dig)

[PCR NcoI-1~8]

Mini-prep of

[Pcons+RBS+NheI-6]

[Pcons+RBS+SalI-1, -7, -11]

Digestion of

[Pcons+RBS+NheI-6](EP)

[Pcons+RBS+SalI-1, -7, -11](EP)

[Pcons+RBS+NcoI]

[Pcons+RBS+Ndel]

PCR of and cultivation of

[Pcons+RBS+NcoI+J61048]

Mini-prep of

[Pcons+RBS+NcoI+J61048-7]Cr

Electrophoresis of

[Pcons+RBS+KphI+J61048-1]Cr

[Pcons+RBS+SalI+J61048-3]Cr

[Pcons+RBS+AseI+J61048 ]Cr

[Pcons+RBS+NdeI+J61048-1]

[Pcons+RBS+HindIII+J61048-1]

[Pcons+RBS+SacI+J61048 -1]

[Pcons+RBS+NcoI+J61048](dig)

[SacI-11]

DNA Sequencing of

[Pcons+RBS+NdeI]-2 Cr

[Pcons+RBS+NcoI]-2 Cr

[Pcons+RBS+NheI]-6 Cr

[Pcons+RBS+SalI-1]Cr

[Pcons+RBS+SalI-11]Cr

[Pcons+RBS+HindIII-1]Cr

[NcoI-12]

[Pcons+RBS+KpnI-1+J61048]Cr

[Pcons+RBS+AseI+J61048]Cr

[Pcons+RBS+SalI-3+J61048]Cr

[Pcons+RBS+HindIII-1+J61048]Cr

[Pcons+RBS+XhoI-2+J61048]Cr

[Pcons+RBS+XhoI-5+J61048]Cr

[Pcons+RBS+NcoI-5+J61048]Cr

[Pcons+RBS+SacI-11-1+J61048]Cr

[Pcons+RBS+NdeI-1+J61048]Cr

[Pcons+RBS+NheI-20+J61048]Cr

[Pcons+RBS+AseI-7+J61048]Cr

[Pcons+RBS+HindIII-7+J61048]Ar

[Pcons+RBS+SacI-16+J61048]Ar

[Pcons+RBS+SalI-17+J61048]Ar

[Pcons+RBS+NdeI-2+J61048]Cr

[Pcons+RBS+NcoI-2+J61048]Cr

[Pcons+RBS+XhoI+J61048]Ar

[Pcons+RBS+KpnI-7+J61048]Ar

[Pcons+RBS+NheI-11+J61048]Ar


10/7

PCR of

[Pcons+RBS+NheI+J61048]

[Pcons+RBS+SalI in PSB3C5]

[Pcons+RBS+SacI in PSB3C5]

Cultivation of

[Pcons+RBS+NheI+J61048-1, -3, -4, -6, -8, -10, -15, -16]

Transformation of

[Pcons+RBS+XhoI-2 +J61048]

[Pcons+RBS+XhoI-5 +J61048-1]

[Pcons+RBS+SacI-11+J61048]

PCR of

[Pcons+RBS+ NdeI in PSB3C5]

[Pcons+RBS+KpnI in PSB3C5]

[Pcons+RBS+AseI in PSB3C5]

[Pcons+RBS+XhoI in PSB3C5]

[Pcons+RBS+NcoI in PSB3C5]

[Pcons+RBS+NheI in PSB3C5]

nine kinds of PBAN

DNA Sequencing of

[Pcons+RBS+NcoI-5+J61048]

10/8

DNA Sequencing of

[PBAN NcoI]

[Pcons+RBS+SalI -1]

[Pcons+RBS+SalI-11]

[Pcons+RBS+NheI-6]

[Pcons+RBS+xhol-2+RBS+BFP+TER]

[Pcons+RBS+SacI-11+RBS+BFP+TER]

[Pcons+RBS+ Nhel-20+RBS+BFP+TER]

Ligation of

Pcons+RBS+NcoI on psb3c5

Pcons+RBS+NcoI on psb3c5

10/9

ALL Nhel-9(10/7)x2

ALL SacI-11(10/7)x1

ALL xhol-2(10/7)x1

Electrophoresis of

Gel 1

Well1 Marker

Well2 Pcons+RBS+NheI-20+RBS+BFP+TER

Well3 Pcons+RBS+Hind-1+RBS+BFP+TER

Well4 Pcons+RBS+KpnI-1+RBS+BFP+TER

Well5 Pcons+RBS+Ndel-1+RBS+BFP+TER

2 Well6 Pcons+RBS+NcoI-5+RBS+BFP+TER

Well7 Pcons+RBS+AseI ALL

Well8 PBAN NcoI-12

Well9 Pcons+RBS+NheI-6

Well10 Pcons+RBS+Hind-1

Well11 Pcons+RBS+NcoI-20

Well12 Pcons+RBS+NcoI-2-2

Well13 Pcons+RBS+Ndel-2-1

Well14 Pcons+RBS+Nedl-2-2

Well15 Pcons+RBS+SacI-1

Well16 Pcons+RBS+SacI-11

Well17 Marker

Gel 2

Well1 marker

Well2 Pcons+RBS+NheI-11

Well3 Pcons+RBS+Hind3

Well4 Pcons+RBS+KpnI-7

Well5 Pcons+RBS+SacI-16

Well6 Pcons+RBS+SalI-12

Well7 Pcons+RBS+Xhol

Transformation of

Pcons+RBS+SacI-3 on psb3c5

Pcons+RBS+KpnI-1 on psb3c5

Pcons+RBS+NcoI-1 on psb3c5

Mini-prep of and sequencing of

ALL NheI-2 x1

ALL SacI-11 x1

ALL Xhol-2 X1

10/10

Transformation of

Pcons+RBS+Ndel on psb3c5

Pcons+RBS+NcoI on psb3c5

Electrophoresis of

Well1 marker

Well2 Pcons+RBS+SacI-3 on psb3c5

Well3 Pcons+RBS+KpnI-1 on psb3c5

Well4 Pcons+RBS+NcoI-1 on psb3c5


10/12

PCR of and Electrophoresis of

Well1 Pcons+RBS+NcoI+psb3c5

Well2 Pcons+RBS+Ndel+psb3c5


None


None


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