Team:NCTU Formosa/Notebook/test7

From 2014.igem.org

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<div class="calendar-note">
<div class="calendar-note">
<div class="single note">1
<div class="single note">1
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10/3
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Cultivation of
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[Pcons+RBS+NdeI]Cr,
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[Pcons+RBS+NcoI]Cr,
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[KpnI-1]Cr, [NdeI-1]Cr,
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[XhoI-1]Cr, [SacI-1]Cr,
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[AseI-1]Cr,
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[ Pcons+RBS+HindIII-1]Cr,
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[ Pcons+RBS+KpnI-4]Cr, [ Pcons+RBS+AseI-5]Cr,
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[ Pcons+RBS+XhoI-1]Cr, [ Pcons+RBS+SacI-1]Cr,
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[ Pcons+RBS+KpnI-1+J61048]Cr, [SalI-4]Cr,
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[Pcons+RBS+SalI-3+J61048]Cr, [HindIII-4]Cr,
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[Pcons+RBS+AseI+J61048]Cr, [NheI], [NcoI]
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<p></p>Electrophoresis of [NdeI](EP), [NcoI](EP), [SacI](EP)
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Transformation of [Pcons+RBS+NdeI]Cr, [Pcons+RBS+SacI]Cr
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Sequencing of [Pcons+RBS+NdeI+J61048-1], [ Pcons+RBS+HindIII+J61048-1], [ Pcons+RBS+SalI+J61048-4], [Pcons+RBS+SacI+J61048-1], the [Pcons+RBS+SacI+J61048-1] was wrong
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10/4
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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
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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]
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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]
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Ligation of [Pcons+RBS+NheI]Kr, [ Pcons+RBS+NcoI]Kr
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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]
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Mini-prep of [Pcons+RBS+NheI-6], [Pcons+RBS+SalI-1, -7, -11]
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Digestion of [Pcons+RBS+NheI-6](EP), [Pcons+RBS+SalI-1, -7, -11](EP), [Pcons+RBS+NcoI], [Pcons+RBS+Ndel]
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PCR and cultivation of [Pcons+RBS+NcoI+J61048]
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Mini-prep of [Pcons+RBS+NcoI+J61048-7]Cr
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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]
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Sequencing of
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[Pcons+RBS+NdeI]-2 Cr
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[Pcons+RBS+NcoI]-2 Cr
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[Pcons+RBS+NheI]-6 Cr
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[Pcons+RBS+SalI-1]Cr
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[Pcons+RBS+SalI-11]Cr
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[Pcons+RBS+HindIII-1]Cr
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[NcoI-12]
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[ Pcons+RBS+KpnI-1+J61048]Cr
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[ Pcons+RBS+AseI+J61048]Cr
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[ Pcons+RBS+SalI-3+J61048]Cr
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[ Pcons+RBS+HindIII-1+J61048]Cr
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[ Pcons+RBS+XhoI-2+J61048]Cr
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[ Pcons+RBS+XhoI-5+J61048]Cr
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[ Pcons+RBS+NcoI-5+J61048]Cr
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[ Pcons+RBS+SacI-11-1+J61048]Cr
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[ Pcons+RBS+NdeI-1+J61048]Cr
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[ Pcons+RBS+NheI-20+J61048]Cr
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[ Pcons+RBS+AseI-7+J61048]Cr
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[ Pcons+RBS+HindIII-7+J61048]Ar
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[ Pcons+RBS+SacI-16+J61048]Ar
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[ Pcons+RBS+SalI-17+J61048]Ar
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[ Pcons+RBS+NdeI-2+J61048]Cr
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[ Pcons+RBS+NcoI-2+J61048]Cr
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[ Pcons+RBS+XhoI+J61048]Ar
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[ Pcons+RBS+KpnI-7+J61048]Ar
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[ Pcons+RBS+NheI-11+J61048]Ar
<br></div>
<br></div>
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<div class="single note"><p>2014/10/8  
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<div class="single note"><p>
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 +
 
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 +
 
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2014/10/8  
Sequencing
Sequencing
1. PBAN NcoI
1. PBAN NcoI

Revision as of 12:58, 13 October 2014

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1 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 Sequencing of [Pcons+RBS+NdeI+J61048-1], [ Pcons+RBS+HindIII+J61048-1], [ Pcons+RBS+SalI+J61048-4], [Pcons+RBS+SacI+J61048-1], the [Pcons+RBS+SacI+J61048-1] was wrong 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 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] 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

2014/10/8 Sequencing 1. PBAN NcoI 2. Pcons+RBS+SalI -1 3. Pcons+RBS+SalI-11 4. Pcons+RBS+NheI-6 5. Pcons+RBS+xhol-2+RBS+BFP+TER 6. Pcons+RBS+SacI-11+RBS+BFP+TER 7. Pcons+RBS+ Nhel-20+RBS+BFP+TER

Ligation Member:Mick Pcons+RBS+NcoI on psb3c5 Pcons+RBS+NcoI on psb3c5

2014/10/9

Cultivation at 12:00

ALL Nhel-9(10/7)x2 ALL SacI-11(10/7)x1 ALL xhol-2(10/7)x1

Electrophoresis The first piece of gel

1. maker 2. Pcons+RBS+NheI-20+RBS+BFP+TER 3. Pcons+RBS+Hind-1+RBS+BFP+TER 4. Pcons+RBS+KpnI-1+RBS+BFP+TER 5. Pcons+RBS+Ndel-1+RBS+BFP+TER 6. Pcons+RBS+NcoI-5+RBS+BFP+TER 7. Pcons+RBS+AseI ALL 8. PBAN NcoI-12 9. Pcons+RBS+NheI-6 10. Pcons+RBS+Hind-1 11. Pcons+RBS+NcoI-20 12. Pcons+RBS+NcoI-2-2 13. Pcons+RBS+Ndel-2-1 14. Pcons+RBS+Nedl-2-2 15. Pcons+RBS+SacI-1 16. Pcons+RBS+SacI-11 17. Maker

The second piece of gel Member: Chris Ho

1. maker 2. Pcons+RBS+NheI-11 3. Pcons+RBS+Hind3 4. Pcons+RBS+KpnI-7 5. Pcons+RBS+SacI-16 6. Pcons+RBS+SalI-12 7. Pcons+RBS+Xhol

Start at 2:15 1 .Pcons+RBS+SacI-3 to psb3c5 2. Pcons+RBS+KpnI-1 to psb3c5 3. Pcons+RBS+NcoI-1 to psb3c5

Mini-prep and sequencing 1. ALL NheI-2 x1 2. ALL SacI-11 x1 3. ALL Xhol-2 X1

2014/10/10 Transform Start at 8:45 Pcons+RBS+Ndel on psb3c5 Pcons+RBS+NcoI on psb3c5

Electrophoresis 1. maker 2. Pcons+RBS+SacI-3 on psb3c5 3. Pcons+RBS+KpnI-1 on psb3c5 4. Pcons+RBS+NcoI-1 on psb3c5


2014/10/10 Transform Start at 8:45 Pcons+RBS+Ndel on psb3c5 Pcons+RBS+NcoI on psb3c5 Electrophoresis 1. maker 2. Pcons+RBS+SacI-3 on psb3c5 3. Pcons+RBS+KpnI-1 on psb3c5 4. Pcons+RBS+NcoI-1 on psb3c5

2014/10/12 PCR and Electrophoresis 1. Pcons+RBS+NcoI+psb3c5 2. Pcons+RBS+Ndel+psb3c5


4
5

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