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
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