Team:NCTU Formosa/Notebook/test7

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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

2014/10/7

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], and nine kinds of PBAN

Sequencing of [Pcons+RBS+NcoI-5+J61048]

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

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