Team:NCTU Formosa/Notebook/test7

From 2014.igem.org

(Difference between revisions)
Line 72: Line 72:
</div>
</div>
<div class="calendar-note">
<div class="calendar-note">
-
<div class="single note">1
+
<div class="single note">
-
10/3
+
<p>
-
<p>Cultivation of  
+
10/3</p><p>
-
[Pcons+RBS+NdeI]Cr,
+
Cultivation of</p><p>
-
[Pcons+RBS+NcoI]Cr,
+
[Pcons+RBS+NdeI]Cr</p><p>
-
[KpnI-1]Cr, [NdeI-1]Cr,
+
[Pcons+RBS+NcoI]Cr</p><p>
-
[XhoI-1]Cr, [SacI-1]Cr,
+
[KpnI-1]Cr</p><p>
-
[AseI-1]Cr,
+
[NdeI-1]Cr</p><p>
-
[ Pcons+RBS+HindIII-1]Cr,
+
[XhoI-1]Cr</p><p>
-
  [ Pcons+RBS+KpnI-4]Cr, [ Pcons+RBS+AseI-5]Cr,
+
[SacI-1]Cr</p><p>
-
[ Pcons+RBS+XhoI-1]Cr, [ Pcons+RBS+SacI-1]Cr,
+
[AseI-1]Cr</p><p>
-
[ Pcons+RBS+KpnI-1+J61048]Cr, [SalI-4]Cr,
+
[ Pcons+RBS+HindIII-1]Cr</p><p>
-
[Pcons+RBS+SalI-3+J61048]Cr, [HindIII-4]Cr,
+
  [ Pcons+RBS+KpnI-4]Cr</p><p>
-
[Pcons+RBS+AseI+J61048]Cr, [NheI], [NcoI]
+
[ Pcons+RBS+AseI-5]Cr</p><p>
-
<p>Electrophoresis of [NdeI](EP), [NcoI](EP), [SacI](EP)
+
[ Pcons+RBS+XhoI-1]Cr</p><p>
-
<p>Transformation of [Pcons+RBS+NdeI]Cr, [Pcons+RBS+SacI]Cr
+
[ Pcons+RBS+SacI-1]Cr</p><p>
-
<p>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
+
[ Pcons+RBS+KpnI-1+J61048]Cr</p><p>
-
10/4  
+
[SalI-4]Cr</p><p>
-
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
+
[Pcons+RBS+SalI-3+J61048]Cr</p><p>
-
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]
+
[HindIII-4]Cr</p><p>
-
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]
+
[Pcons+RBS+AseI+J61048]Cr</p><p>
-
Ligation of [Pcons+RBS+NheI]Kr, [ Pcons+RBS+NcoI]Kr
+
[NheI]</p><p>
-
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]
+
[NcoI]</p><p>
-
Mini-prep of [Pcons+RBS+NheI-6], [Pcons+RBS+SalI-1, -7, -11]
+
Electrophoresis of</p><p>
-
Digestion of [Pcons+RBS+NheI-6](EP), [Pcons+RBS+SalI-1, -7, -11](EP), [Pcons+RBS+NcoI], [Pcons+RBS+Ndel]
+
[NdeI](EP)</p><p>
-
PCR and cultivation of [Pcons+RBS+NcoI+J61048]
+
[NcoI](EP)</p><p>
-
Mini-prep of [Pcons+RBS+NcoI+J61048-7]Cr
+
[SacI](EP)</p><p>
-
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]
+
Transformation of</p><p>
-
Sequencing of  
+
[Pcons+RBS+NdeI]Cr</p><p>
-
[Pcons+RBS+NdeI]-2 Cr
+
[Pcons+RBS+SacI]Cr</p><p>
-
[Pcons+RBS+NcoI]-2 Cr
+
Sequencing of</p><p>
-
[Pcons+RBS+NheI]-6 Cr
+
[Pcons+RBS+NdeI+J61048-1]</p><p>
-
[Pcons+RBS+SalI-1]Cr
+
[ Pcons+RBS+HindIII+J61048-1]</p><p>
-
[Pcons+RBS+SalI-11]Cr
+
[ Pcons+RBS+SalI+J61048-4]</p><p>
-
[Pcons+RBS+HindIII-1]Cr
+
[Pcons+RBS+SacI+J61048-1]</p><p>
-
[NcoI-12]
+
[Pcons+RBS+SacI+J61048-1](fail)</p><p>
-
[ Pcons+RBS+KpnI-1+J61048]Cr
+
10/4</p><p>
-
[ Pcons+RBS+AseI+J61048]Cr
+
Digestion of</p><p>
-
[ Pcons+RBS+SalI-3+J61048]Cr
+
[Pcons+RBS+XhoI+J61048-2, -4, -5] </p><p>
-
[ Pcons+RBS+HindIII-1+J61048]Cr
+
[Pcons+RBS+SacI+J61048-10, -11]</p><p>
-
[ Pcons+RBS+XhoI-2+J61048]Cr
+
[Pcons+RBS+NheI+J61048-20]</p><p>
-
[ Pcons+RBS+XhoI-5+J61048]Cr
+
[Pcons+RBS+NheI]Cr</p><p>
-
[ Pcons+RBS+NcoI-5+J61048]Cr
+
[Pcons+RBS+NcoI]Cr</p><p>
-
[ Pcons+RBS+SacI-11-1+J61048]Cr
+
Cultivation of </p><p>
-
[ Pcons+RBS+NdeI-1+J61048]Cr
+
[Pcons+RBS+NheI-1, -2, -4, -6, -10, -12]</p><p>
-
[ Pcons+RBS+NheI-20+J61048]Cr
+
[Pcons+RBS+SalI-1, -3, -6, -7, -11, -12] </p><p>
-
[ Pcons+RBS+AseI-7+J61048]Cr
+
Cultivation of [Pcons+RBS+NcoI+J61048-1~8]</p><p>
-
[ Pcons+RBS+HindIII-7+J61048]Ar
+
PCR of</p><p>
-
[ Pcons+RBS+SacI-16+J61048]Ar
+
[Pcons+RBS+NheI-1, -2, -4, -6, -10, -12]</p><p>
-
[ Pcons+RBS+SalI-17+J61048]Ar
+
[Pcons+RBS+SacI-1, -3, -6, -7, -11, -12]</p><p>
-
[ Pcons+RBS+NdeI-2+J61048]Cr
+
[Pcons+RBS+NcoI+J61048-1~8]</p><p>
-
[ Pcons+RBS+NcoI-2+J61048]Cr
+
Ligation of </p><p>
-
[ Pcons+RBS+XhoI+J61048]Ar
+
[Pcons+RBS+NheI]Kr</p><p>
-
[ Pcons+RBS+KpnI-7+J61048]Ar
+
[ Pcons+RBS+NcoI]Kr</p><p>
-
[ Pcons+RBS+NheI-11+J61048]Ar
+
Electrophoresis of </p><p>
 +
[XhoI-2, -4, -5](dig)</p><p>
 +
[SacI-10, -11](dig)</p><p>
 +
[NheI-20](dig)</p><p>
 +
[Pcons+RBS+NdeI](dig)</p><p>
 +
[Pcons+RBS+NcoI](dig)</p><p>
 +
[PCR NcoI-1~8]</p><p>
 +
Mini-prep of</p><p>
 +
[Pcons+RBS+NheI-6]</p><p>
 +
[Pcons+RBS+SalI-1, -7, -11]</p><p>
 +
Digestion of </p><p>
 +
[Pcons+RBS+NheI-6](EP)</p><p>
 +
[Pcons+RBS+SalI-1, -7, -11](EP)</p><p>
 +
[Pcons+RBS+NcoI]</p><p>
 +
[Pcons+RBS+Ndel]</p><p>
 +
PCR and cultivation of </p><p>
 +
[Pcons+RBS+NcoI+J61048]</p><p>
 +
 
 +
Mini-prep of</p><p>
 +
[Pcons+RBS+NcoI+J61048-7]Cr</p><p>
 +
Electrophoresis of </p><p>
 +
[Pcons+RBS+KphI+J61048-1]Cr</p><p>
 +
[ Pcons+RBS+SalI+J61048-3]Cr</p><p>
 +
[ Pcons+RBS+AseI+J61048 ]Cr</p><p>
 +
[Pcons+RBS+NdeI+J61048-1]</p><p>
 +
[ Pcons+RBS+HindIII+J61048-1]</p><p>
 +
[ Pcons+RBS+SacI+J61048 -1]</p><p>
 +
[Pcons+RBS+NcoI+J61048](dig)</p><p>
 +
[SacI-11]</p><p>
 +
Sequencing of </p><p>
 +
[Pcons+RBS+NdeI]-2 Cr</p><p>
 +
[Pcons+RBS+NcoI]-2 Cr</p><p>
 +
[Pcons+RBS+NheI]-6 Cr</p><p>
 +
[Pcons+RBS+SalI-1]Cr</p><p>
 +
[Pcons+RBS+SalI-11]Cr</p><p>
 +
[Pcons+RBS+HindIII-1]Cr</p><p>
 +
[NcoI-12]</p><p>
 +
[ Pcons+RBS+KpnI-1+J61048]Cr</p><p>
 +
[ Pcons+RBS+AseI+J61048]Cr</p><p>
 +
[ Pcons+RBS+SalI-3+J61048]Cr</p><p>
 +
[ Pcons+RBS+HindIII-1+J61048]Cr</p><p>
 +
[ Pcons+RBS+XhoI-2+J61048]Cr</p><p>
 +
[ Pcons+RBS+XhoI-5+J61048]Cr</p><p>
 +
[ Pcons+RBS+NcoI-5+J61048]Cr</p><p>
 +
[ Pcons+RBS+SacI-11-1+J61048]Cr</p><p>
 +
[ Pcons+RBS+NdeI-1+J61048]Cr</p><p>
 +
[ Pcons+RBS+NheI-20+J61048]Cr</p><p>
 +
[ Pcons+RBS+AseI-7+J61048]Cr</p><p>
 +
[ Pcons+RBS+HindIII-7+J61048]Ar</p><p>
 +
[ Pcons+RBS+SacI-16+J61048]Ar</p><p>
 +
[ Pcons+RBS+SalI-17+J61048]Ar</p><p>
 +
[ Pcons+RBS+NdeI-2+J61048]Cr</p><p>
 +
[ Pcons+RBS+NcoI-2+J61048]Cr</p><p>
 +
[ Pcons+RBS+XhoI+J61048]Ar</p><p>
 +
[ Pcons+RBS+KpnI-7+J61048]Ar</p><p>
 +
[ Pcons+RBS+NheI-11+J61048]Ar</p>
<br></div>
<br></div>
-
<div class="single note"><p>
+
<div class="single note">
-
 
+
-
 
+
-
2014/10/7
+
-
<p>PCR of [ Pcons+RBS+NheI+J61048], [ Pcons+RBS+SalI in PSB3C5], [Pcons+RBS+SacI in PSB3C5]
+
-
<p>Cultivation of [Pcons+RBS+NheI+J61048-1, -3, -4, -6, -8, -10, -15, -16]
+
-
<p>Transformation of [Pcons+RBS+XhoI-2 +J61048], [Pcons+RBS+XhoI-5 +J61048-1], [Pcons+RBS+SacI-11+J61048],
+
-
<p>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
+
-
<p>Sequencing of  [Pcons+RBS+NcoI-5+J61048]
+
-
</p>
+
-
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
+
-
</p>
+
<p>
<p>
-
Ligation
+
10/7</p><p>
-
Member:Mick
+
PCR of</p><p>
-
Pcons+RBS+NcoI on psb3c5
+
[ Pcons+RBS+NheI+J61048]</p><p>
-
Pcons+RBS+NcoI on psb3c5
+
[ Pcons+RBS+SalI in PSB3C5]</p><p>
-
</p><p>
+
[Pcons+RBS+SacI in PSB3C5]</p><p>
-
2014/10/9
+
Cultivation of </p><p>
-
</p><p>
+
[Pcons+RBS+NheI+J61048-1, -3, -4, -6, -8, -10, -15, -16]</p><p>
-
Cultivation at 12:00
+
Transformation of </p><p>
-
</p>
+
[Pcons+RBS+XhoI-2 +J61048]</p><p>
-
ALL Nhel-9(10/7)x2
+
[Pcons+RBS+XhoI-5 +J61048-1]</p><p>
-
ALL SacI-11(10/7)x1
+
[Pcons+RBS+SacI-11+J61048]</p><p>
-
ALL xhol-2(10/7)x1
+
PCR of</p><p>
-
<p>
+
  [Pcons+RBS+ NdeI  in PSB3C5]</p><p>
-
Electrophoresis
+
[Pcons+RBS+KpnI in PSB3C5]</p><p>
-
The first piece of gel
+
[Pcons+RBS+AseI in PSB3C5]</p><p>
-
</p>
+
[Pcons+RBS+XhoI in PSB3C5]</p><p>
-
1. maker
+
[Pcons+RBS+NcoI in PSB3C5]</p><p>
-
2. Pcons+RBS+NheI-20+RBS+BFP+TER
+
[Pcons+RBS+NheI in PSB3C5]</p><p>
-
3. Pcons+RBS+Hind-1+RBS+BFP+TER
+
nine kinds of PBAN</p><p>
-
4. Pcons+RBS+KpnI-1+RBS+BFP+TER
+
Sequencing of</p><p>
-
5. Pcons+RBS+Ndel-1+RBS+BFP+TER
+
  [Pcons+RBS+NcoI-5+J61048]</p><p>
-
6. Pcons+RBS+NcoI-5+RBS+BFP+TER
+
10/8 </p><p>
-
7. Pcons+RBS+AseI ALL
+
Sequencing of</p><p>
-
8. PBAN NcoI-12
+
[PBAN NcoI]</p><p>
-
9. Pcons+RBS+NheI-6
+
[Pcons+RBS+SalI -1]</p><p>
-
10. Pcons+RBS+Hind-1
+
[Pcons+RBS+SalI-11]</p><p>
-
11. Pcons+RBS+NcoI-20
+
[Pcons+RBS+NheI-6]</p><p>
-
12. Pcons+RBS+NcoI-2-2
+
[Pcons+RBS+xhol-2+RBS+BFP+TER]</p><p>
-
13. Pcons+RBS+Ndel-2-1
+
[Pcons+RBS+SacI-11+RBS+BFP+TER]</p><p>
-
14. Pcons+RBS+Nedl-2-2
+
[Pcons+RBS+ Nhel-20+RBS+BFP+TER]</p>
-
15. Pcons+RBS+SacI-1
+
-
16. Pcons+RBS+SacI-11
+
-
17. Maker
+
<p>
<p>
-
The second piece of gel
+
Ligation of</p><p>
-
Member: Chris Ho
+
Pcons+RBS+NcoI on psb3c5</p><p>
-
</p><p>
+
Pcons+RBS+NcoI on psb3c5</p><p>
-
1. maker
+
10/9</p><p>
-
2. Pcons+RBS+NheI-11
+
ALL Nhel-9(10/7)x2</p><p>
-
3. Pcons+RBS+Hind3
+
ALL SacI-11(10/7)x1</p><p>
-
4. Pcons+RBS+KpnI-7
+
ALL xhol-2(10/7)x1</p><p>
-
5. Pcons+RBS+SacI-16
+
Electrophoresis of</p><p>
-
6. Pcons+RBS+SalI-12
+
gel 1</p><p>
-
7. Pcons+RBS+Xhol
+
Well1 Marker</p><p>
 +
Well2 Pcons+RBS+NheI-20+RBS+BFP+TER</p><p>
 +
Well3 Pcons+RBS+Hind-1+RBS+BFP+TER</p><p>
 +
Well4 Pcons+RBS+KpnI-1+RBS+BFP+TER </p><p>
 +
Well5 Pcons+RBS+Ndel-1+RBS+BFP+TER</p><p>
 +
Well6 Pcons+RBS+NcoI-5+RBS+BFP+TER</p><p>
 +
Well7 Pcons+RBS+AseI ALL</p><p>
 +
Well8 PBAN NcoI-12</p><p>
 +
Well9 Pcons+RBS+NheI-6</p><p>
 +
Well10 Pcons+RBS+Hind-1</p><p>
 +
Well11 Pcons+RBS+NcoI-20</p><p>
 +
Well12 Pcons+RBS+NcoI-2-2</p><p>
 +
Well13 Pcons+RBS+Ndel-2-1</p><p>
 +
Well14 Pcons+RBS+Nedl-2-2</p><p>
 +
Well15 Pcons+RBS+SacI-1</p><p>
 +
Well16 Pcons+RBS+SacI-11</p><p>
 +
Well17 Marker</p><p>
 +
gel 2</p><p>
 +
Well1 marker</p><p>
 +
Well2 Pcons+RBS+NheI-11</p><p>
 +
Well3 Pcons+RBS+Hind3</p><p>
 +
Well4 Pcons+RBS+KpnI-7</p><p>
 +
Well5 Pcons+RBS+SacI-16</p><p>
 +
Well6 Pcons+RBS+SalI-12</p><p>
 +
Well7 Pcons+RBS+Xhol</p><p>
 +
Transformation of</p><p>
 +
Pcons+RBS+SacI-3 on psb3c5</p><p>
 +
Pcons+RBS+KpnI-1 on psb3c5</p><p>
 +
Pcons+RBS+NcoI-1 on psb3c5
</p><p>
</p><p>
 +
Mini-prep and sequencing</p><p>
 +
ALL NheI-2 x1</p><p>
 +
ALL SacI-11 x1</p><p>
 +
ALL Xhol-2 X1</p><p>
 +
10/10</p><p>
 +
Transformation of </p><p>
 +
Pcons+RBS+Ndel on psb3c5</p><p>
 +
Pcons+RBS+NcoI on psb3c5</p><p>
 +
Electrophoresis of </p><p>
-
Start at 2:15
+
Well1 marker</p><p>
-
1 .Pcons+RBS+SacI-3 to psb3c5
+
Well2 Pcons+RBS+SacI-3 on psb3c5</p><p>
-
2. Pcons+RBS+KpnI-1 to psb3c5
+
Well3 Pcons+RBS+KpnI-1 on psb3c5</p><p>
-
3. Pcons+RBS+NcoI-1 to psb3c5
+
Well4 Pcons+RBS+NcoI-1 on psb3c5</p><p>
-
</p><p>
+
-
 
+
-
Mini-prep and sequencing
+
-
1. ALL NheI-2 x1
+
-
2. ALL SacI-11 x1
+
-
3. ALL Xhol-2 X1
+
-
</p><p>
+
-
2014/10/10
+
-
Transform
+
-
Start at 8:45
+
-
Pcons+RBS+Ndel on psb3c5
+
-
Pcons+RBS+NcoI on psb3c5
+
-
</p><p>
+
-
Electrophoresis
+
-
 
+
-
1. maker
+
-
2. Pcons+RBS+SacI-3 on psb3c5
+
-
3. Pcons+RBS+KpnI-1 on psb3c5
+
-
4. Pcons+RBS+NcoI-1 on psb3c5
+
-
</p>
+
<br></div>
<br></div>
-
<div class="single note"><p>
+
<div class="single note">
-
2014/10/10
+
<p>
-
Transform
+
10/10</p><p>
-
Start at 8:45
+
Transformation of</p><p>
-
Pcons+RBS+Ndel on psb3c5
+
Pcons+RBS+Ndel on psb3c5</p><p>
-
Pcons+RBS+NcoI on psb3c5
+
Pcons+RBS+NcoI on psb3c5</p><p>
-
Electrophoresis
+
Electrophoresis of</p><p>
-
1. maker
+
Well1 marker</p><p>
-
2. Pcons+RBS+SacI-3 on psb3c5
+
Well2 Pcons+RBS+SacI-3 on psb3c5</p><p>
-
3. Pcons+RBS+KpnI-1 on psb3c5
+
Well3 Pcons+RBS+KpnI-1 on psb3c5</p><p>
-
4. Pcons+RBS+NcoI-1 on psb3c5
+
Well4 Pcons+RBS+NcoI-1 on psb3c5</p><p>
-
</p><p>
+
10/12</p><p>
-
2014/10/12
+
PCR and Electrophoresis of</p><p>
-
PCR and Electrophoresis
+
-
1. Pcons+RBS+NcoI+psb3c5
+
Well1 Pcons+RBS+NcoI+psb3c5</p><p>
-
2. Pcons+RBS+Ndel+psb3c5
+
Well2 Pcons+RBS+Ndel+psb3c5</p>
-
</p>
+
<br></div>
<br></div>
<div class="single note">4
<div class="single note">4

Revision as of 15:26, 13 October 2014

  • Sunday
  • Monday
  • Tuesday
  • Wednesday
  • Thursday
  • Friday
  • Saturday
  • 　
  • 　
  • 　
  • 1
  • 2
  • 3
  • 4
  • 5
  • 6
  • 7
  • 8
  • 9
  • 10
  • 11
  • 12
  • 13
  • 14
  • 15
  • 16
  • 17
  • 18
  • 19
  • 20
  • 21
  • 22
  • 23
  • 24
  • 25
  • 26
  • 27
  • 28
  • 29
  • 30
  • 31
  • 　

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]

[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 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


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

Sequencing of

[Pcons+RBS+NcoI-5+J61048]

10/8

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

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 and sequencing

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


10/10

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 and Electrophoresis of

Well1 Pcons+RBS+NcoI+psb3c5

Well2 Pcons+RBS+Ndel+psb3c5


4
5

Retrieved from "http://2014.igem.org/Team:NCTU_Formosa/Notebook/test7"