"
Team:NCTU Formosa/Notebook/test6
From 2014.igem.org
| Line 69: | Line 69: | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 2014/9/4 | + | 2014/9/4 |
Cultivation of: | Cultivation of: | ||
Pcons+RBS+AseI PCR product-1,2,3,4,7,8 Ar | Pcons+RBS+AseI PCR product-1,2,3,4,7,8 Ar | ||
| Line 86: | Line 86: | ||
[PBAN-HindIII] Kr*2 (EP), [PBAN-NheI]Kr*2 (EP), [PBAN-KpnI-HF]Kr*2 (EP),[23101+B0034+PBAN (HindIII)]Ar(ES),[23101+B0034+PBAN (NheI)]Ar(ES),[23101+B0034+PBAN (HindIII)]Ar(EP)*2,[23101+B0034+PBAN (NheI)]Ar(EP),[23101+B0034+PBAN (KpnI-HF)]Ar(EP)*2,[23101+B0034+PBAN (KpnI-HF)]Ar PCR*3. | [PBAN-HindIII] Kr*2 (EP), [PBAN-NheI]Kr*2 (EP), [PBAN-KpnI-HF]Kr*2 (EP),[23101+B0034+PBAN (HindIII)]Ar(ES),[23101+B0034+PBAN (NheI)]Ar(ES),[23101+B0034+PBAN (HindIII)]Ar(EP)*2,[23101+B0034+PBAN (NheI)]Ar(EP),[23101+B0034+PBAN (KpnI-HF)]Ar(EP)*2,[23101+B0034+PBAN (KpnI-HF)]Ar PCR*3. | ||
</p><p> | </p><p> | ||
| - | 2014/9/5 | + | 2014/9/5 |
Electrophoresis: | Electrophoresis: | ||
Well 1 Pcons+RBS+AseI--1 (EP) Ar | Well 1 Pcons+RBS+AseI--1 (EP) Ar | ||
| Line 128: | Line 128: | ||
<p> | <p> | ||
| - | 2014/9/9 | + | 2014/9/9 |
Cultivation of: | Cultivation of: | ||
Pcons+RBS+SacI PCR product-16 Ar | Pcons+RBS+SacI PCR product-16 Ar | ||
| Line 136: | Line 136: | ||
XhoI+PSB1K3 (EP) | XhoI+PSB1K3 (EP) | ||
</p><p> | </p><p> | ||
| - | 2014/9/10 | + | 2014/9/10 |
Mini prep of: | Mini prep of: | ||
RBS+BFP+Ter-16 Ar | RBS+BFP+Ter-16 Ar | ||
| Line 161: | Line 161: | ||
Well8：XhoI-7 dig(XP) Kr | Well8：XhoI-7 dig(XP) Kr | ||
</p><p> | </p><p> | ||
| - | 2014/9/11 | + | 2014/9/11 |
Digestion of: | Digestion of: | ||
| - | Pcons+RBS+PBAN(ES)( | + | Pcons+RBS+PBAN(ES)(8 kind except of XhoI) |
RBS+BFP+Ter(XP) | RBS+BFP+Ter(XP) | ||
Ligation of: | Ligation of: | ||
| Line 188: | Line 188: | ||
Pcons+RBS+HindIII dig(EP) | Pcons+RBS+HindIII dig(EP) | ||
Pcons+RBS+NheI dig(EP) | Pcons+RBS+NheI dig(EP) | ||
| - | Pcons+RBS+NcoI dig(EP)-->9/ | + | Pcons+RBS+NcoI dig(EP)-->9/11 mini |
Pcons+RBS+NdeI dig(EP) | Pcons+RBS+NdeI dig(EP) | ||
Pcons+RBS+SacI dig(EP) | Pcons+RBS+SacI dig(EP) | ||
| Line 199: | Line 199: | ||
| - | 2014/9/12 | + | 2014/9/12 |
Transformation of: | Transformation of: | ||
Pcons+RBS+NheI+RBS+BFP+Ter Kr | Pcons+RBS+NheI+RBS+BFP+Ter Kr | ||
| Line 223: | Line 223: | ||
</p> | </p> | ||
<p> | <p> | ||
| - | 2014/9/13 | + | 2014/9/13 |
Ligation of: | Ligation of: | ||
Pcons+RBS+AseI Kr | Pcons+RBS+AseI Kr | ||
Revision as of 14:21, 10 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
2014/9/4 Cultivation of: Pcons+RBS+AseI PCR product-1,2,3,4,7,8 Ar XhoI+PSB1K3 PCR product-1,2 Kr PSB1A3 Ar Cultivation plate test: Ar plate and Kr plate Digestion of: [PBAN-HindIII] Kr*2 (EP), [PBAN-NheI]Kr*2 (EP), [PBAN-KpnI-HF]Kr*2 (EP) 23101+B0034+PBAN (HindIII)]Ar(ES),[23101+B0034+PBAN (NheI)]Ar(ES) PCR of: [J23101+B0034+KpnI-HF] Ar*3 Ligation of: [J23101+B0034+KpnI-HF+PSB1C3] Electrophoresis of: [PBAN-HindIII] Kr*2 (EP), [PBAN-NheI]Kr*2 (EP), [PBAN-KpnI-HF]Kr*2 (EP),[23101+B0034+PBAN (HindIII)]Ar(ES),[23101+B0034+PBAN (NheI)]Ar(ES),[23101+B0034+PBAN (HindIII)]Ar(EP)*2,[23101+B0034+PBAN (NheI)]Ar(EP),[23101+B0034+PBAN (KpnI-HF)]Ar(EP)*2,[23101+B0034+PBAN (KpnI-HF)]Ar PCR*3.
2014/9/5 Electrophoresis: Well 1 Pcons+RBS+AseI--1 (EP) Ar Well 2 Pcons+RBS+AseI--2 (EP) Ar Well 3 Pcons+RBS+AseI--3 (EP) Ar Well 4 Pcons+RBS+AseI--4 (EP) Ar Well 5 Pcons+RBS+AseI--7 (EP) Ar （DNA sequencing） Well 6 Pcons+RBS+AseI--8 (EP) Ar Well 7 Pcons+RBS+NcoI--2 (EP) Cr （DNA sequencing） Well 8 Pcons+RBS+Ndel--2 (EP) Cr （DNA sequencing） Well 9 1 Kb marker Well 10 Pcons+RBS+Ndel--13 (EP) Cr Well 11 SacI--3 (XP) Kr Well 12 SacI--4.1 (XP) Kr Well 13 SacI--4.2 (XP) Kr Well 14 XhoI--7 (EP) Kr （DNA sequencing） Well 15 XhoI--8 (EP) Kr（DNA sequencing） Well 16 SalI--3 (XP) Kr Mini prep of: Pcons+RBS+KpnI-HF-7 Pcons+RBS+KpnI-HF-8 Pcons+RBS+KpnI-HF-9 Digestion of: Pcons+RBS+KpnI-HF-7 (EP) Pcons+RBS+KpnI-HF-8 (EP) Pcons+RBS+KpnI-HF-9 (EP) Cultivation of: Pcons+RBS+KpnI-HF Ar PCR product-7,8,9 Ar Ligation of: [J23101+B0034+HindIII+B0034+PSB1K3] Kr [J23101+B0034+HindIII+J61048+PSB1K3] Kr [J23101+B0034+NheI+B0034+PSB1K3] Kr [J23101+B0034+NheI+J61048+PSB1K3] Kr Electrophoresis of digested product: [Pcons+RBS+KpnI-HF]Ar (EP)*3
2014/9/9 Cultivation of: Pcons+RBS+SacI PCR product-16 Ar Pcons+RBS+SaII PCR product-17 Ar RBS+BFP+Ter PCR product-16,17,29,30 Ar Digestion of: XhoI+PSB1K3 (EP)
2014/9/10 Mini prep of: RBS+BFP+Ter-16 Ar RBS+BFP+Ter-17 Ar RBS+BFP+Ter-29 Ar RBS+BFP+Ter-30 Ar Pcons+RBS+Sac-16 Ar Pcons+RBS+Sal-17 Ar Digestion of: RBS+BFP+Ter-16 Ar(EP) RBS+BFP+Ter-17 Ar(EP) RBS+BFP+Ter-29 Ar(EP) RBS+BFP+Ter-30 Ar(EP) Pcons+RBS+SacI-16 Ar(EP) Pcons+RBS+SalI-17 Ar(EP) Electrophoresis: Well1：Marker Well2：Pcons+RBS+SacI-16 dig(EP) Well3：Pcons+RBS+SalI-17 dig(EP) Well4：RBS+BFP+Ter-16 dig(EP) Well5：RBS+BFP+Ter-17 dig(EP) Well6：RBS+BFP+Ter-29 dig(EP) Well7：RBS+BFP+Ter-30 dig(EP) Well8：XhoI-7 dig(XP) Kr
2014/9/11 Digestion of: Pcons+RBS+PBAN(ES)(8 kind except of XhoI) RBS+BFP+Ter(XP) Ligation of: Pcons+RBS+XhoI-7 Ar Cultivation of: Pcons+RBS+AseI-7 Ar Pcons+RBS+SacI-16 Ar Pcons+RBS+SalI-17 Ar Pcons+RBS+NcoI-2 Ar Pcons+RBS+NdeI-2 Ar RBS+BFP+Ter-2 Ar RBS+BFP+Ter-2 Ar SalI-3 Kr Ligation of: Pcons+RBS+XhoI-7 Ar DNA sequencing of: Pcons+RBS+SaeI Ar 8ul VF2 Pcons+RBS+SalI Ar 8ul VF2 RBS+BFP+Ter-29 Ar 8ul VF2 + 8ul VR RBS+BFP+Ter-30 Ar 5ul VF2 + 5ul VR Digestion of: Pcons+RBS+HindIII dig(EP) Pcons+RBS+NheI dig(EP) Pcons+RBS+NcoI dig(EP)-->9/11 mini Pcons+RBS+NdeI dig(EP) Pcons+RBS+SacI dig(EP) Pcons+RBS+SalI dig(EP) Pcons+RBS+AseI dig(EP) Pcons+RBS+KpnI dig(EP) Ligation of: Pcons+RBS+BFP+Ter-30 Kr
2014/9/12 Transformation of: Pcons+RBS+NheI+RBS+BFP+Ter Kr Pcons+RBS+NdeI+RBS+BFP+Ter Kr Pcons+RBS+HindIII+RBS+BFP+Ter Kr Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr Pcons+RBS+AseI+RBS+BFP+Ter Kr Pcons+RBS+SacI+RBS+BFP+Ter Kr Pcons+RBS+NcoI+RBS+BFP+Ter Kr Pcons+RBS+SalI+RBS+BFP+Ter Kr Pcons+RBS+BFP+Ter Kr Pcons+RBS+XhoI Ar Electrophoresis: Well1：Marker Well2：Pcons+RBS+NdeI dig(EP) Cr Well3：Pcons+RBS+NcoI dig(EP) Cr Well4：Pcons+RBS+SacI dig(EP) Ar Well5：Pcons+RBS+SalI dig(EP) Ar Well6：Pcons+RBS+AseI dig(EP) Ar Well7：Pcons+RBS+HindIII dig(EP) Ar Well8：Pcons+RBS+NheI dig(EP) Ar Well9：Pcons+RBS+KpnI dig(EP) Ar
2014/9/13 Ligation of: Pcons+RBS+AseI Kr Pcons+RBS+NdeI Kr Pcons+RBS+SalI Kr Pcons+RBS+SacI Kr PCR of: Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8 Pcons+RBS+NheI+RBS+BFP+Ter Kr x8 Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8 Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8 Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8 Pcons+RBS+AseI+RBS+BFP+Ter Kr x8 Pcons+RBS+Xhol Kr x8 Pcons+RBS+SalI+RBS+BFP+Ter Kr x16 Pcons+RBS+SacI+RBS+BFP+Ter Kr x16 Electrophoresis: Gel 1 Pcons+RBS+SalI+RBS+BFP+Ter Kr x16 Gel 2 Pcons+RBS+SacI+RBS+BFP+Ter Kr x16 Gel 3 Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8 Pcons+RBS+NheI+RBS+BFP+Ter Kr x8 Gel 4 Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8 Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8 Gel 5 Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8 Pcons+RBS+AseI+RBS+BFP+Ter Kr x8 Gel 6 Pcons+RBS+Xhol Kr x8 psb1K3 dig(EP)
