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Team:NCTU Formosa/Notebook/test6
From 2014.igem.org
| Line 103: | Line 103: | ||
9/5</p> | 9/5</p> | ||
<p> | <p> | ||
| - | Electrophoresis:</p> | + | Electrophoresis:</p><p> |
| - | Well 1 Pcons+RBS+AseI--1 (EP) Ar | + | Well 1 Pcons+RBS+AseI--1 (EP) Ar</p><p> |
| - | Well 2 Pcons+RBS+AseI--2 (EP) Ar | + | Well 2 Pcons+RBS+AseI--2 (EP) Ar</p><p> |
| - | Well 3 Pcons+RBS+AseI--3 (EP) Ar | + | Well 3 Pcons+RBS+AseI--3 (EP) Ar</p><p> |
| - | Well 4 Pcons+RBS+AseI--4 (EP) Ar | + | Well 4 Pcons+RBS+AseI--4 (EP) Ar</p><p> |
| - | Well 5 Pcons+RBS+AseI--7 (EP) Ar （DNA sequencing） | + | Well 5 Pcons+RBS+AseI--7 (EP) Ar （DNA sequencing）</p><p> |
| - | Well 6 Pcons+RBS+AseI--8 (EP) Ar | + | Well 6 Pcons+RBS+AseI--8 (EP) Ar</p><p> |
| - | Well 7 Pcons+RBS+NcoI--2 (EP) Cr （DNA sequencing） | + | Well 7 Pcons+RBS+NcoI--2 (EP) Cr （DNA sequencing）</p><p> |
| - | Well 8 Pcons+RBS+Ndel--2 (EP) Cr | + | Well 8 Pcons+RBS+Ndel--2 (EP) Cr （DNA sequencing）</p><p> |
| - | Well 9 1 Kb marker | + | Well 9 1 Kb marker</p><p> |
| - | Well 10 Pcons+RBS+Ndel--13 (EP) Cr | + | Well 10 Pcons+RBS+Ndel--13 (EP) Cr</p><p> |
| - | Well 11 SacI--3 (XP) Kr | + | Well 11 SacI--3 (XP) Kr</p><p> |
| - | Well 12 SacI--4.1 (XP) Kr | + | Well 12 SacI--4.1 (XP) Kr</p><p> |
| - | Well 13 SacI--4.2 (XP) Kr | + | Well 13 SacI--4.2 (XP) Kr</p><p> |
| - | Well 14 XhoI--7 (EP) Kr （DNA sequencing） | + | Well 14 XhoI--7 (EP) Kr （DNA sequencing）</p><p> |
| - | Well 15 XhoI--8 (EP) Kr（DNA sequencing） | + | Well 15 XhoI--8 (EP) Kr（DNA sequencing）</p><p> |
| - | Well 16 SalI--3 (XP) Kr | + | Well 16 SalI--3 (XP) Kr</p><p> |
| - | <p> | + | Mini prep of:</p><p> |
| - | Mini prep of:</p> | + | Pcons+RBS+KpnI-HF-7</p><p> |
| - | Pcons+RBS+KpnI-HF-7 | + | Pcons+RBS+KpnI-HF-8</p><p> |
| - | Pcons+RBS+KpnI-HF-8 | + | Pcons+RBS+KpnI-HF-9</p><p> |
| - | Pcons+RBS+KpnI-HF-9 | + | Digestion of:</p><p> |
| - | <p> | + | Pcons+RBS+KpnI-HF-7 (EP)</p><p> |
| - | Digestion of:</p> | + | Pcons+RBS+KpnI-HF-8 (EP)</p><p> |
| - | Pcons+RBS+KpnI-HF-7 (EP) | + | Pcons+RBS+KpnI-HF-9 (EP)</p><p> |
| - | + | Cultivation of:</p><p> | |
| - | Pcons+RBS+KpnI-HF-9 | + | Pcons+RBS+KpnI-HF Ar PCR product-7,8,9 Ar</p><p> |
| - | + | Ligation of: </p><p> | |
| - | <p>Cultivation of:</p> | + | [J23101+B0034+HindIII+B0034+PSB1K3] Kr</p><p> |
| - | Pcons+RBS+KpnI-HF Ar PCR product-7,8,9 Ar | + | [J23101+B0034+HindIII+J61048+PSB1K3] Kr</p><p> |
| - | Ligation of: | + | [J23101+B0034+NheI+B0034+PSB1K3] Kr</p><p> |
| - | [J23101+B0034+HindIII+B0034+PSB1K3] Kr | + | [J23101+B0034+NheI+J61048+PSB1K3] Kr</p><p> |
| - | [J23101+B0034+HindIII+J61048+PSB1K3] Kr | + | Electrophoresis of digested product:</p><p> |
| - | [J23101+B0034+NheI+B0034+PSB1K3] Kr | + | [Pcons+RBS+KpnI-HF]Ar (EP)*3 </p><p> |
| - | [J23101+B0034+NheI+J61048+PSB1K3] Kr | + | |
| - | Electrophoresis of digested product: | + | |
| - | [Pcons+RBS+KpnI-HF]Ar (EP)*3 | + | |
</p> | </p> | ||
| Line 145: | Line 142: | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| + | 9/9</p><p> | ||
| + | Cultivation of:</p><p> | ||
| + | Pcons+RBS+SacI PCR product-16 Ar</p><p> | ||
| + | Pcons+RBS+SaII PCR product-17 Ar</p><p> | ||
| + | RBS+BFP+Ter PCR product-16,17,29,30 Ar</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | XhoI+PSB1K3 (EP)</p><p> | ||
| + | 9/10</p><p> | ||
| + | Mini prep of:</p><p> | ||
| + | RBS+BFP+Ter-16 Ar</p><p> | ||
| + | RBS+BFP+Ter-17 Ar</p><p> | ||
| + | RBS+BFP+Ter-29 Ar</p><p> | ||
| + | RBS+BFP+Ter-30 Ar</p><p> | ||
| + | Pcons+RBS+Sac-16 Ar</p><p> | ||
| + | Pcons+RBS+Sal-17 Ar</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | RBS+BFP+Ter-16 Ar(EP)</p><p> | ||
| + | RBS+BFP+Ter-17 Ar(EP)</p><p> | ||
| + | RBS+BFP+Ter-29 Ar(EP)</p><p> | ||
| + | RBS+BFP+Ter-30 Ar(EP)</p><p> | ||
| + | Pcons+RBS+SacI-16 Ar(EP)</p><p> | ||
| + | Pcons+RBS+SalI-17 Ar(EP)</p><p> | ||
| + | Electrophoresis:</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：Pcons+RBS+SacI-16 dig(EP)</p><p> | ||
| + | Well3：Pcons+RBS+SalI-17 dig(EP)</p><p> | ||
| + | Well4：RBS+BFP+Ter-16 dig(EP)</p><p> | ||
| + | Well5：RBS+BFP+Ter-17 dig(EP)</p><p> | ||
| + | Well6：RBS+BFP+Ter-29 dig(EP)</p><p> | ||
| + | Well7：RBS+BFP+Ter-30 dig(EP)</p><p> | ||
| + | Well8：XhoI-7 dig(XP) Kr</p><p> | ||
| + | 9/11</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | Pcons+RBS+PBAN(ES)(8 kind except of XhoI)</p><p> | ||
| + | RBS+BFP+Ter(XP)</p><p> | ||
| + | Ligation of:</p><p> | ||
| + | Pcons+RBS+XhoI-7 Ar</p><p> | ||
| + | Cultivation of:</p><p> | ||
| + | Pcons+RBS+AseI-7 Ar</p><p> | ||
| + | Pcons+RBS+SacI-16 Ar</p><p> | ||
| + | Pcons+RBS+SalI-17 Ar</p><p> | ||
| + | Pcons+RBS+NcoI-2 Ar</p><p> | ||
| + | Pcons+RBS+NdeI-2 Ar</p><p> | ||
| + | RBS+BFP+Ter-2 Ar</p><p> | ||
| + | RBS+BFP+Ter-2 Ar</p><p> | ||
| + | SalI-3 Kr</p><p> | ||
| + | Ligation of:</p><p> | ||
| + | Pcons+RBS+XhoI-7 Ar</p><p> | ||
| - | + | DNA sequencing of:</p><p> | |
| - | + | Pcons+RBS+SaeI Ar 8ul VF2</p><p> | |
| - | + | Pcons+RBS+SalI Ar 8ul VF2</p><p> | |
| - | + | RBS+BFP+Ter-29 Ar 8ul VF2 + 8ul VR</p><p> | |
| - | + | RBS+BFP+Ter-30 Ar 5ul VF2 + 5ul VR</p><p> | |
| - | + | Digestion of:</p><p> | |
| - | + | Pcons+RBS+HindIII dig(EP)</p><p> | |
| - | </p><p> | + | Pcons+RBS+NheI dig(EP)</p><p> |
| - | + | Pcons+RBS+NcoI dig(EP)-->9/11 mini</p><p> | |
| - | + | Pcons+RBS+NdeI dig(EP)</p><p> | |
| - | + | Pcons+RBS+SacI dig(EP)</p><p> | |
| - | + | Pcons+RBS+SalI dig(EP)</p><p> | |
| - | + | Pcons+RBS+AseI dig(EP)</p><p> | |
| - | + | Pcons+RBS+KpnI dig(EP)</p><p> | |
| - | Pcons+RBS+ | + | Ligation of: </p><p> |
| - | + | Pcons+RBS+BFP+Ter-30 Kr</p><p> | |
| - | + | 9/12</p><p> | |
| - | + | Transformation of:</p><p> | |
| - | + | Pcons+RBS+NheI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+NdeI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+HindIII+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+AseI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+SacI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+NcoI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+SalI+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+BFP+Ter Kr</p><p> | |
| - | + | Pcons+RBS+XhoI Ar</p><p> | |
| - | + | Electrophoresis:</p><p> | |
| - | + | Well1：Marker</p><p> | |
| - | + | Well2：Pcons+RBS+NdeI dig(EP) Cr</p><p> | |
| - | + | Well3：Pcons+RBS+NcoI dig(EP) Cr</p><p> | |
| - | </p><p> | + | Well4：Pcons+RBS+SacI dig(EP) Ar</p><p> |
| - | + | Well5：Pcons+RBS+SalI dig(EP) Ar</p><p> | |
| - | + | Well6：Pcons+RBS+AseI dig(EP) Ar</p><p> | |
| - | + | Well7：Pcons+RBS+HindIII dig(EP) Ar</p><p> | |
| - | + | Well8：Pcons+RBS+NheI dig(EP) Ar</p><p> | |
| - | + | Well9：Pcons+RBS+KpnI dig(EP) Ar</p> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | 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 | + | |
| - | </p><p> | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | 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 | + | |
| - | </p> | + | |
<p> | <p> | ||
| - | 2014/9/13 | + | 2014/9/13</p><p> |
| - | Ligation of: | + | Ligation of:</p><p> |
| - | Pcons+RBS+AseI Kr | + | Pcons+RBS+AseI Kr</p><p> |
| - | Pcons+RBS+NdeI Kr | + | Pcons+RBS+NdeI Kr</p><p> |
| - | Pcons+RBS+SalI Kr | + | Pcons+RBS+SalI Kr</p><p> |
| - | Pcons+RBS+SacI Kr | + | Pcons+RBS+SacI Kr</p><p> |
| - | PCR of: | + | PCR of:</p><p> |
| - | Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+NheI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NheI+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+AseI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+AseI+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+Xhol Kr x8 | + | Pcons+RBS+Xhol Kr x8</p><p> |
| - | Pcons+RBS+SalI+RBS+BFP+Ter Kr x16 | + | Pcons+RBS+SalI+RBS+BFP+Ter Kr x16</p><p> |
| - | Pcons+RBS+SacI+RBS+BFP+Ter Kr x16 | + | Pcons+RBS+SacI+RBS+BFP+Ter Kr x16</p><p> |
| - | Electrophoresis: | + | Electrophoresis:</p><p> |
| - | Gel 1 | + | Gel 1</p><p> |
| - | Pcons+RBS+SalI+RBS+BFP+Ter Kr x16 | + | Pcons+RBS+SalI+RBS+BFP+Ter Kr x16</p><p> |
| - | Gel 2 | + | Gel 2</p><p> |
| - | Pcons+RBS+SacI+RBS+BFP+Ter Kr x16 | + | Pcons+RBS+SacI+RBS+BFP+Ter Kr x16</p><p> |
| - | Gel 3 | + | Gel 3</p><p> |
| - | Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+NheI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NheI+RBS+BFP+Ter Kr x8</p><p> |
| - | Gel 4 | + | Gel 4</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8</p><p> |
| - | Gel 5 | + | Gel 5</p><p> |
| - | Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8</p><p> |
| - | Pcons+RBS+AseI+RBS+BFP+Ter Kr x8 | + | Pcons+RBS+AseI+RBS+BFP+Ter Kr x8</p><p> |
| - | Gel 6 | + | Gel 6</p><p> |
| - | Pcons+RBS+Xhol Kr x8 | + | Pcons+RBS+Xhol Kr x8</p><p> |
| - | psb1K3 dig(EP) | + | psb1K3 dig(EP)</p> |
| - | </p> | + | |
<br></div> | <br></div> | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | 2014/9/16 | + | 2014/9/16</p><p> |
| - | Transformation of | + | Transformation of</p><p> |
| - | psb3K3 *2 | + | psb3K3 *2</p><p> |
| - | psb3C3 | + | psb3C3</p><p> |
| - | Digestion of: | + | Digestion of:</p><p> |
| - | psb3K3 dig(EP) | + | psb3K3 dig(EP)</p><p> |
| - | psb1C3 dig(EP) | + | psb1C3 dig(EP)</p><p> |
| - | Cultivation of: | + | Cultivation of:</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter x8 | + | Pcons+RBS+PBAN+RBS+BFP+Ter x8</p> |
| - | </p> | + | |
<p> | <p> | ||
| - | + | 9/17</p><p> | |
| - | Mini prep of: | + | Mini prep of:</p><p> |
| - | PBAN：XhoI | + | PBAN：XhoI</p><p> |
| - | SalI | + | SalI</p><p> |
| - | SacI | + | SacI</p><p> |
| - | KpnI | + | KpnI</p><p> |
| - | Ndel | + | Ndel</p><p> |
| - | AseI | + | AseI</p><p> |
| - | NcoI | + | NcoI</p><p> |
| - | NheI | + | NheI</p><p> |
| - | HindIII (fail) | + | HindIII (fail)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4 | + | Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4</p><p> |
| - | Ndel-8 | + | Ndel-8</p><p> |
| - | HindIII-4 (fail) | + | HindIII-4 (fail)</p><p> |
| - | SacI-9(fail) | + | SacI-9(fail)</p><p> |
| - | KpnI-1 | + | KpnI-1</p><p> |
| - | NcoI-2 | + | NcoI-2</p><p> |
| - | NheI-1 | + | NheI-1</p><p> |
| - | Digestion of: | + | Digestion of:</p><p> |
| - | PBAN：XhoI | + | PBAN：XhoI</p><p> |
| - | SalI | + | SalI</p><p> |
| - | SacI | + | SacI</p><p> |
| - | KpnI | + | KpnI</p><p> |
| - | Ndel | + | Ndel</p><p> |
| - | AseI | + | AseI</p><p> |
| - | NcoI | + | NcoI</p><p> |
| - | AseI | + | AseI</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4 | + | Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4</p><p> |
| - | Ndel-8 | + | Ndel-8</p><p> |
| - | KpnI-1 | + | KpnI-1</p><p> |
| - | NcoI-2 | + | NcoI-2</p><p> |
| - | NheI-1 | + | NheI-1</p><p> |
| - | Electrophoresis: | + | Electrophoresis:</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (Ndel) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (Ndel)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (NcoI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (NcoI)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (SacI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (SalI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (NheI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (NheI)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (KpnI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (KpnI)</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (AseI) | + | Pcons+RBS+PBAN+RBS+BFP+Ter (AseI)</p><p> |
| - | PBAN AseI | + | PBAN AseI</p><p> |
| - | PBAN Ndel | + | PBAN Ndel</p><p> |
| - | PBAN KpnI | + | PBAN KpnI</p><p> |
| - | PBAN SacI | + | PBAN SacI</p><p> |
| - | PBAN SalI | + | PBAN SalI</p><p> |
| - | PBAN XhoI | + | PBAN XhoI</p><p> |
| - | Pcons+RBS+XhoI-1 dig(EP) | + | Pcons+RBS+XhoI-1 dig(EP)</p><p> |
| - | Pcons+RBS+Xhol-1 dig(ES) | + | Pcons+RBS+Xhol-1 dig(ES)</p><p> |
| - | psb1C3 dig(ES) | + | psb1C3 dig(ES)</p><p> |
| - | Cultivation of: | + | Cultivation of:</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)-4.12 | + | Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)-4.12</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)-6 | + | Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)-6</p><p> |
| - | Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)-9 | + | Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)-9</p><p> |
| - | Digestion of: | + | Digestion of:</p><p> |
| - | B0034 Kr dig(ES) | + | B0034 Kr dig(ES)</p><p> |
| - | Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP) | + | Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP) | + | Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | NcoI Kr dig(EP) | + | NcoI Kr dig(EP)</p><p> |
| - | Ndel Kr dig(EP) | + | Ndel Kr dig(EP)</p><p> |
| - | SalI Kr dig(EP) | + | SalI Kr dig(EP)</p><p> |
| - | + | 9/18</p><p> | |
| - | </p><p> | + | Electrophoresis:</p><p> |
| - | + | Well 1：Marker</p><p> | |
| - | Electrophoresis: | + | Well2：B0034 Kr dig(ES)</p><p> |
| - | + | Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)</p><p> | |
| - | Well2：B0034 Kr dig(ES) | + | Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP) | + | Well5：NcoI Kr dig(EP)</p><p> |
| - | Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP) | + | Well6：Ndel Kr dig(EP)</p><p> |
| - | Well5：NcoI Kr dig(EP) | + | Well7：SalI Kr dig(EP)</p><p> |
| - | Well6：Ndel Kr dig(EP) | + | Electrophoresis:</p><p> |
| - | Well7：SalI Kr dig(EP) | + | Well1：Marker</p><p> |
| - | Electrophoresis: | + | Well2：B0034 Kr dig(ES)</p><p> |
| - | Well1：Marker | + | Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Well2：B0034 Kr dig(ES) | + | Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP) | + | Well5：NcoI Kr dig(EP)</p><p> |
| - | Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP) | + | Well6：Ndel Kr dig(EP)</p><p> |
| - | Well5：NcoI Kr dig(EP) | + | Well7：SalI Kr dig(EP)</p><p> |
| - | Well6：Ndel Kr dig(EP) | + | Mini prep:</p><p> |
| - | Well7：SalI Kr dig(EP) | + | Pcons+RBS+HindIII+RBS+BFP+Ter-6</p><p> |
| - | Mini prep: | + | Pcons+RBS+SalI+RBS+BFP+Ter-9</p><p> |
| - | Pcons+RBS+HindIII+RBS+BFP+Ter-6 | + | Pcons+RBS+SacI+RBS+BFP+Ter-4</p><p> |
| - | Pcons+RBS+SalI+RBS+BFP+Ter-9 | + | Pcons+RBS+SacI+RBS+BFP+Ter-11</p><p> |
| - | Pcons+RBS+SacI+RBS+BFP+Ter-4 | + | Ligation of:</p><p> |
| - | Pcons+RBS+SacI+RBS+BFP+Ter-11 | + | Pcons+RBS+AseI+RBS+BFP+Ter Cr</p><p> |
| - | Ligation of: | + | Pcons+RBS+NdeI+RBS+BFP+Ter Cr</p><p> |
| - | Pcons+RBS+AseI+RBS+BFP+Ter Cr | + | NheI Cr</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter Cr | + | NcoI Cr</p> |
| - | NheI Cr | + | |
| - | NcoI Cr | + | |
| - | + | ||
| - | </p> | + | |
<p> | <p> | ||
| + | 9/20</p><p> | ||
| + | Transformation of:</p><p> | ||
| + | AseI mini Cr</p><p> | ||
| - | + | Electrophoresis:</p><p> | |
| - | + | Well1：Marker</p><p> | |
| - | + | Well2：HindIII-2 Kr dig(EP)</p><p> | |
| - | + | Well3：SacI Kr dig(EP)</p><p> | |
| - | + | Well4：Pcons+RBS+HindIII-6+RBS+BFP+Ter Kr dig(EP)</p><p> | |
| - | Electrophoresis: | + | Well5：Pcons+RBS+SacI-11+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Well1：Marker | + | Well6：Pcons+RBS+SacI-9+RBS+BFP+Ter Kr dig(EP)</p><p> |
| - | Well2：HindIII-2 Kr dig(EP) | + | Well7：RBS+BFP+Ter Ar dig(XP)</p><p> |
| - | Well3：SacI Kr dig(EP) | + | Well8：Pcons+RBS+NheI+RBS+BFP+Ter Kr dig(EP)</p> |
| - | Well4：Pcons+RBS+HindIII-6+RBS+BFP+Ter Kr dig(EP) | + | |
| - | Well5：Pcons+RBS+SacI-11+RBS+BFP+Ter Kr dig(EP) | + | |
| - | Well6：Pcons+RBS+SacI-9+RBS+BFP+Ter Kr dig(EP) | + | |
| - | Well7：RBS+BFP+Ter Ar dig(XP) | + | |
| - | Well8：Pcons+RBS+NheI+RBS+BFP+Ter Kr dig(EP) | + | |
| - | </p> | + | |
<br></div> | <br></div> | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | + | 9/21</p><p> | |
| - | Electrophoresis of PCR products: | + | Electrophoresis of PCR products:</p><p> |
| - | Gel 1 | + | Gel 1</p><p> |
| - | Well1：Marker | + | Well1：Marker</p><p> |
| - | Well2：3H1 | + | Well2：3H1</p><p> |
| - | Well3：3H2 | + | Well3：3H2</p><p> |
| - | Well4：3H3 | + | Well4：3H3</p><p> |
| - | Well5：3H4 | + | Well5：3H4</p><p> |
| - | Well6：3H5 | + | Well6：3H5</p><p> |
| - | Well7：3H6 | + | Well7：3H6</p><p> |
| - | Well8：Marker | + | Well8：Marker</p><p> |
| - | Well9：6K1 | + | Well9：6K1</p><p> |
| - | Well10：6K2 | + | Well10：6K2</p><p> |
| - | Well11：6K3 | + | Well11：6K3</p><p> |
| - | Well12：6K4 | + | Well12：6K4</p><p> |
| - | Well13：6K5 | + | Well13：6K5</p><p> |
| - | Well14：6K6 | + | Well14：6K6</p><p> |
| - | Well15：3Sl1 | + | Well15：3Sl1</p><p> |
| - | Well16：3Sl2 | + | Well16：3Sl2</p><p> |
| - | Well17：3Sl3 | + | Well17：3Sl3</p><p> |
| - | Gel 2 | + | Gel 2</p><p> |
| - | Well1：Marker | + | Well1：Marker</p><p> |
| - | Well2：6Sl4 | + | Well2：6Sl4</p><p> |
| - | Well3：6Sl5 | + | Well3：6Sl5</p><p> |
| - | Well4：3Nh1 | + | Well4：3Nh1</p><p> |
| - | Well5：3Nh2 | + | Well5：3Nh2</p><p> |
| - | Well6：3Nh3 | + | Well6：3Nh3</p><p> |
| - | Well7：3Nh4 | + | Well7：3Nh4</p><p> |
| - | Well8：Marker | + | Well8：Marker</p><p> |
| - | Well9：3Nh5 | + | Well9：3Nh5</p><p> |
| - | Well10：3Nh6 | + | Well10：3Nh6</p><p> |
| - | Well11：6X1 | + | Well11：6X1</p><p> |
| - | Well12：6X2 | + | Well12：6X2</p><p> |
| - | Well13：6X3 | + | Well13：6X3</p><p> |
| - | Well14：6X4 | + | Well14：6X4</p><p> |
| - | Well15：6X5 | + | Well15：6X5</p><p> |
| - | Well16：6X6 | + | Well16：6X6</p><p> |
| - | Well17：1K1 | + | Well17：1K1</p><p> |
| - | Gel 3 | + | Gel 3</p><p> |
| - | Well1：Marker | + | Well1：Marker</p><p> |
| - | Well2：1K2 | + | Well2：1K2</p><p> |
| - | Well3：1K3 | + | Well3：1K3</p><p> |
| - | Well4：1K4 | + | Well4：1K4</p><p> |
| - | Well5：1K5 | + | Well5：1K5</p><p> |
| - | Well6：1K6 | + | Well6：1K6</p><p> |
| - | Well7：1X1 | + | Well7：1X1</p><p> |
| - | Well8：Marker | + | Well8：Marker</p><p> |
| - | Well9：1X2 | + | Well9：1X2</p><p> |
| - | Well10：1X3 | + | Well10：1X3</p><p> |
| - | Well11：1X4 | + | Well11：1X4</p><p> |
| - | Well12：1X5 | + | Well12：1X5</p><p> |
| - | Well13：1X6 | + | Well13：1X6</p><p> |
| - | Well14：3K1 | + | Well14：3K1</p><p> |
| - | Well15：3K2 | + | Well15：3K2</p><p> |
| - | Well16：3K3 | + | Well16：3K3</p><p> |
| - | Well17：3K4 | + | Well17：3K4</p><p> |
| - | Gel 4 | + | Gel 4</p><p> |
| - | Well1：Marker | + | Well1：Marker</p><p> |
| - | Well2：3K5 | + | Well2：3K5</p><p> |
| - | Well3：3K6 | + | Well3：3K6</p><p> |
| - | Well4：3Sc1 | + | Well4：3Sc1</p><p> |
| - | Well5：3Sc2 | + | Well5：3Sc2</p><p> |
| - | Well6：3Sc3 | + | Well6：3Sc3</p><p> |
| - | Well7：3Sc4 | + | Well7：3Sc4</p><p> |
| - | Well8：Marker | + | Well8：Marker</p><p> |
| - | Well9：3Sc5 | + | Well9：3Sc5</p><p> |
| - | Well10：3Sc6 | + | Well10：3Sc6</p><p> |
| - | Well11：1Sc1 | + | Well11：1Sc1</p><p> |
| - | Well12：1Sc2 | + | Well12：1Sc2</p><p> |
| - | Well13：1Sc3 | + | Well13：1Sc3</p><p> |
| - | Well14：1Sc4 | + | Well14：1Sc4</p><p> |
| - | Well15：1Sc5 | + | Well15：1Sc5</p><p> |
| - | Well16：1Sc6 | + | Well16：1Sc6</p><p> |
| - | Well17：1Nc1 | + | Well17：1Nc1</p><p> |
| + | Gel 5</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：1Nc2</p><p> | ||
| + | Well3：1Nc3</p><p> | ||
| + | Well4：1Nc4</p><p> | ||
| + | Well5：1Nc5</p><p> | ||
| + | Well6：1Nc6</p><p> | ||
| + | Well7：1A1</p><p> | ||
| + | Well8：Marker</p><p> | ||
| + | Well9：1A2</p><p> | ||
| + | Well10：1A3</p><p> | ||
| + | Well11：1A4</p><p> | ||
| + | Well12：1A5</p><p> | ||
| + | Well13：1A6</p><p> | ||
| + | Well14：1Nd1</p><p> | ||
| + | Well15：1Nd2</p><p> | ||
| + | Well16：1Nd3</p><p> | ||
| + | Well17：1Nd4</p><p> | ||
| + | Gel 6</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：1Nd5</p><p> | ||
| + | Well3：1Nd6</p><p> | ||
| + | Well4：3X1</p><p> | ||
| + | Well5：3X2</p><p> | ||
| + | Well6：3X3</p><p> | ||
| + | Well7：3X4</p><p> | ||
| + | Well8：Marker</p><p> | ||
| + | Well9：3X5</p><p> | ||
| + | Well10：3X6</p><p> | ||
| + | Well11：3A1</p><p> | ||
| + | Well12：3A2</p><p> | ||
| + | Well13：3A3</p><p> | ||
| + | Well14：3A4</p><p> | ||
| + | Well15：3A5</p><p> | ||
| + | Well16：3A6</p><p> | ||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| + | Cultivation of:</p><p> | ||
| + | KpnI-1</p><p> | ||
| + | NdeI-1</p><p> | ||
| + | XhoI-1</p><p> | ||
| + | SacI-1</p><p> | ||
| + | AseI-1</p><p> | ||
| + | Pcons+RBS+HindIII-1</p><p> | ||
| + | Pcons+RBS+KpnI-4</p><p> | ||
| + | Pcons+RBS+NheI-4</p><p> | ||
| + | Pcons+RBS+SalI-5</p><p> | ||
| + | Pcons+RBS+AseI-5</p><p> | ||
| + | Pcons+RBS+XhoI-1</p><p> | ||
| + | Pcons+RBS+SacI-1</p><p> | ||
| + | Pcons+RBS+KpnI-1+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+XhoI-6+RBS+BFP+Ter</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | SalI Kr dig(EP)</p><p> | ||
| + | Ligation of:</p><p> | ||
| + | NdeI dig(EP)+C backbone</p><p> | ||
| + | 9/22</p><p> | ||
| + | Transformation of:</p><p> | ||
| + | NheI Cr</p><p> | ||
| + | Pcons+RBS+NheI+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+Ndel+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+AseI+RBS+BFP+Ter Cr</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | HindIII(EP)</p><p> | ||
| + | Cultivation of:</p><p> | ||
| + | SacI-3 Kr</p><p> | ||
| + | NcoI-7 Cr</p><p> | ||
| + | NcoI-8 Cr</p><p> | ||
| + | Pcons+RBS+SacI-2</p><p> | ||
| + | Pcons+RBS+XhoI-5+RBS+BFP+Ter</p><p> | ||
| + | pbs3C5</p><p> | ||
| + | Ligation of:</p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+HindIII+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+SacI+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+SalI+RBS+BFP+Ter Cr</p><p> | ||
| + | 9/23</p><p> | ||
| + | Electrophoresis:</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：KpnI-1 Cr dig(EP)</p><p> | ||
| + | Well3：NdeI-1 Cr dig(EP)</p><p> | ||
| + | Well4：XhoI-1 Cr dig(EP)</p><p> | ||
| + | Well5：SacI-1 Cr dig(EP)</p><p> | ||
| + | Well6：AseI-1 Cr dig(EP)</p><p> | ||
| + | Well7：Pcons+RBS+HindIII-1 dig(EP)</p><p> | ||
| + | Well8：Pcons+RBS+KpnI-4 dig(EP)</p><p> | ||
| + | Well9：Pcons+RBS+NheI-4 dig(EP)</p><p> | ||
| + | Well10：Pcons+RBS+AseI-5 dig(EP)</p><p> | ||
| + | Well11：Pcons+RBS+XhoI-1 dig(EP)</p><p> | ||
| + | Well12：Pcons+RBS+SacI-1 dig(EP)</p><p> | ||
| + | Well13：Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)</p><p> | ||
| + | Well14：SalI Kr dig(EP) 9/21</p><p> | ||
| + | Well15：SalI Kr dig(EP) 9/19</p><p> | ||
| + | DNA sequencing of:</p><p> | ||
| + | KpnI-1 Cr dig(EP)</p><p> | ||
| + | NdeI-1 Cr dig(EP)</p><p> | ||
| + | XhoI-1 Cr dig(EP)</p><p> | ||
| + | SacI-1 Cr dig(EP)</p><p> | ||
| + | AseI-1 Cr dig(EP)</p><p> | ||
| + | Pcons+RBS+HindIII-1 dig(EP)</p><p> | ||
| + | Pcons+RBS+KpnI-4 dig(EP)</p><p> | ||
| + | Pcons+RBS+NheI-4 dig(EP)(fail)</p><p> | ||
| + | Pcons+RBS+AseI-5 dig(EP)</p><p> | ||
| + | Pcons+RBS+XhoI-1 dig(EP)</p><p> | ||
| + | Pcons+RBS+SacI-1 dig(EP)</p><p> | ||
| + | Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)</p><p> | ||
| + | Transformation of:</p><p> | ||
| + | Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+SalI+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+SacI+RBS+BFP+Ter</p><p> | ||
| + | 9/24</p><p> | ||
| + | Cultivation of:</p><p> | ||
| + | Pcons+RBS+NheI+RBS+BFP+Ter Cr</p><p> | ||
| + | Pcons+RBS+AseI+RBS+BFP+Ter Cr </p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter Cr </p><p> | ||
| + | Pcons+RBS+NdeI+RBS+BFP+Ter Cr </p><p> | ||
| + | NheI Cr </p><p> | ||
| + | psb3C5 Cr </p><p> | ||
| + | Cultivation of:</p><p> | ||
| + | Pcons+RBS+NheI+RBS+BFP+Ter Cr </p><p> | ||
| + | Pcons+RBS+AseI+RBS+BFP+Ter Cr </p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter Cr </p><p> | ||
| + | Pcons+RBS+NdeI+RBS+BFP+Ter Cr </p><p> | ||
| + | NheI Cr </p><p> | ||
| + | psb3C5 Cr </p><p> | ||
| + | Electrophoresis:</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：SalI Kr dig(EP)</p><p> | ||
| + | Well3：Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)</p><p> | ||
| + | Well4：Pcons+RBS+SacI Cr dig(EP)</p><p> | ||
| + | Well5：NcoI Cr dig(EP)</p><p> | ||
| + | Digestion of:</p><p> | ||
| + | Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)</p><p> | ||
| + | Pcons+RBS+SacI-2 Cr dig(EP)</p><p> | ||
| + | NcoI-8 Cr dig(EP)</p><p> | ||
| + | SalI-3 Kr dig(EP)</p><p> | ||
| + | Ligation of:</p><p> | ||
| + | HindIII+psb1C3</p><p> | ||
| + | PCR of:</p><p> | ||
| + | SacI x8</p><p> | ||
| + | SalI x5</p><p> | ||
| + | HindIII x2</p><p> | ||
| + | NheI x8</p><p> | ||
| + | Mini prep and digestion of:</p><p> | ||
| + | Pcons+RBS+AseI+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+NheI+RBS+BFP+Ter </p><p> | ||
| + | Pcons+RBS+NdeI+RBS+BFP+Ter</p><p> | ||
| + | Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> | ||
| + | psb3C5 </p><p> | ||
| + | NheI</p><p> | ||
| + | 9/25</p><p> | ||
| + | Electrophoresis and DNAsequencing:</p><p> | ||
| + | Well1：Marker</p><p> | ||
| + | Well2：NheI (fail)</p><p> | ||
| + | Well3：Pcons+RBS+AseI+RBS+BFP+Ter</p><p> | ||
| + | Well4：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> | ||
| + | Well5：Pcons+RBS+NdeI+RBS+BFP+Ter (fail)</p><p> | ||
| - | + | Transformation of:</p><p> | |
| - | + | HindIII</p><p> | |
| - | + | SalI</p><p> | |
| - | + | Pcons+RBS+SacI+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+SalI+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | Pcons+RBS+ | + | |
| - | + | ||
| - | + | ||
| - | Pcons+RBS+ | + | |
| - | Pcons+RBS+ | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | </p><p> | + | PCR of:</p><p> |
| + | NcoI x5(9.10.11.12.13)</p><p> | ||
| + | Pcons+RBS+SacI x5(7.8.9.10.11)</p><p> | ||
| - | + | Cultivation of:</p><p> | |
| - | + | NheI</p><p> | |
| - | NheI | + | Pcons+RBS+SacI-2(9/22)</p><p> |
| - | Pcons+RBS+ | + | Pcons+RBS+NcoI-8(9/22)</p><p> |
| - | Pcons+RBS+NcoI | + | Pcons+RBS+NheI-4</p><p> |
| - | + | Pcons+RBS+NdeI+RBS+BFP+Ter x2</p><p> | |
| - | + | Pcons+RBS+NheI+RBS+BFP+Ter x2</p><p> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | Pcons+RBS+ | + | |
| - | Pcons+RBS+ | + | |
| - | + | ||
| - | + | ||
| - | Pcons+RBS+ | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | </p><p> | + | 9/26</p><p> |
| - | + | Mini prep and digestion(EP) of:</p><p> | |
| - | + | NcoI</p><p> | |
| - | + | NheI</p><p> | |
| - | + | Pcons+RBS+SacI-2</p><p> | |
| - | + | Pcons+RBS+NheI-4</p><p> | |
| - | + | Pcons+RBS+NdeI-1+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+NdeI-2+RBS+BFP+Ter</p><p> | |
| - | + | Electrophoresis:</p><p> | |
| - | + | Well1：Marker</p><p> | |
| - | + | Well2：Pcons+RBS+NdeI-1+RBS+BFP+Ter</p><p> | |
| - | + | Well3：Pcons+RBS+NdeI-2+RBS+BFP+Ter</p><p> | |
| - | + | Well4：NcoI</p><p> | |
| - | + | Well5：NheI</p><p> | |
| - | + | Well6：Pcons+RBS+NheI-4</p><p> | |
| - | + | Well7：Pcons+RBS+SacI-2</p><p> | |
| - | + | Cultivation of:</p><p> | |
| - | + | psb3C5 x2</p><p> | |
| - | + | HindIII-3</p><p> | |
| - | + | HindIII-4</p><p> | |
| - | + | SalI-3</p><p> | |
| - | + | SalI-4</p><p> | |
| - | + | Pcons+RBS+HindIII-3+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+HindIII-4+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+SalI-3+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+SalI-4+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+SacI-3+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+SacI-4+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+NcoI-3+RBS+BFP+Ter</p><p> | |
| - | + | Pcons+RBS+NcoI-4+RBS+BFP+Ter</p> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | </p><p> | + | |
| - | + | ||
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| - | + | ||
| - | </p><p> | + | |
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| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | Pcons+RBS+SacI-2 | + | |
| - | + | ||
| - | Pcons+RBS+NheI-4 | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | </p><p> | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | Pcons+RBS+NdeI-1+RBS+BFP+Ter | + | |
| - | Pcons+RBS+NdeI-2+RBS+BFP+Ter | + | |
| - | Electrophoresis: | + | |
| - | Well1：Marker | + | |
| - | Well2：Pcons+RBS+NdeI-1+RBS+BFP+Ter | + | |
| - | Well3：Pcons+RBS+NdeI-2+RBS+BFP+Ter | + | |
| - | Well4：NcoI | + | |
| - | Well5：NheI | + | |
| - | Well6：Pcons+RBS+NheI-4 | + | |
| - | Well7：Pcons+RBS+SacI-2 | + | |
| - | Cultivation of: | + | |
| - | psb3C5 x2 | + | |
| - | HindIII-3 | + | |
| - | HindIII-4 | + | |
| - | SalI-3 | + | |
| - | SalI-4 | + | |
| - | Pcons+RBS+HindIII-3+RBS+BFP+Ter | + | |
| - | Pcons+RBS+HindIII-4+RBS+BFP+Ter | + | |
| - | Pcons+RBS+SalI-3+RBS+BFP+Ter | + | |
| - | Pcons+RBS+SalI-4+RBS+BFP+Ter | + | |
| - | Pcons+RBS+SacI-3+RBS+BFP+Ter | + | |
| - | Pcons+RBS+SacI-4+RBS+BFP+Ter | + | |
| - | Pcons+RBS+NcoI-3+RBS+BFP+Ter | + | |
| - | Pcons+RBS+NcoI-4+RBS+BFP+Ter | + | |
| - | </p> | + | |
<br></div> | <br></div> | ||
<div class="single note"> | <div class="single note"> | ||
<p> | <p> | ||
| - | + | 9/28</p><p> | |
| - | Electrophoresis: | + | Electrophoresis:</p><p> |
| - | Well1：HindIII-3 | + | Well1：HindIII-3</p><p> |
| - | Well2：HindIII-4 | + | Well2：HindIII-4</p><p> |
| - | Well3：SalI-3 | + | Well3：SalI-3</p><p> |
| - | Well4：SalI-4 | + | Well4：SalI-4</p><p> |
| - | Well5：Pcons+RBS+SacI-3+RBS+BFP+Ter | + | Well5：Pcons+RBS+SacI-3+RBS+BFP+Ter</p><p> |
| - | Well6：Marker | + | Well6：Marker</p><p> |
| - | Well7：Pcons+RBS+NcoI-3+RBS+BFP+Ter | + | Well7：Pcons+RBS+NcoI-3+RBS+BFP+Ter</p><p> |
| - | Well8：Pcons+RBS+NcoI-4+RBS+BFP+Ter | + | Well8：Pcons+RBS+NcoI-4+RBS+BFP+Ter</p><p> |
| - | Well9：Pcons+RBS+SalI-3+RBS+BFP+Ter | + | Well9：Pcons+RBS+SalI-3+RBS+BFP+Ter</p><p> |
| - | Well10：Pcons+RBS+SalI-4+RBS+BFP+Ter | + | Well10：Pcons+RBS+SalI-4+RBS+BFP+Ter</p><p> |
| - | Well11：psb3C5 | + | Well11：psb3C5</p><p> |
| - | </p><p> | + | 9/29</p><p> |
| - | + | Electrophoresis:</p><p> | |
| - | Electrophoresis: | + | Well1：Marker</p><p> |
| - | Well1：Marker | + | Well2：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well2：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well3：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well3：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well4：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well4：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well5：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well5：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well6：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well6：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well7：Pcons+RBS+HindIII+RBS+BFP+Ter</p><p> |
| - | Well7：Pcons+RBS+HindIII+RBS+BFP+Ter | + | Well8：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well8：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well9：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well9：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well10：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well10：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well11：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well11：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well12：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well12：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well13：Pcons+RBS+NcoI+RBS+BFP+Ter</p><p> |
| - | Well13：Pcons+RBS+NcoI+RBS+BFP+Ter | + | Well14：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well14：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well15：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well15：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well16：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well16：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well17：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well17：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well18：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well18：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well19：Pcons+RBS+SalI+RBS+BFP+Ter</p><p> |
| - | Well19：Pcons+RBS+SalI+RBS+BFP+Ter | + | Well20：psb3C5 dig</p><p> |
| - | Well20：psb3C5 dig | + | Well21：Marker</p><p> |
| - | Well21：Marker | + | Ligation of:</p><p> |
| - | Ligation of: | + | Pcons+RBS+KpnI + pbs3C5</p><p> |
| - | Pcons+RBS+KpnI + pbs3C5 | + | Pcons+RBS+SacI + pbs3C5</p><p> |
| - | Pcons+RBS+SacI + pbs3C5 | + | Pcons+RBS+AseI + pbs3C5</p><p> |
| - | Pcons+RBS+AseI + pbs3C5 | + | Pcons+RBS+HindIII + pbs3C5</p><p> |
| - | Pcons+RBS+HindIII + pbs3C5 | + | Pcons+RBS+NheI + pbs3C5</p><p> |
| - | Pcons+RBS+NheI + pbs3C5 | + | Pcons+RBS+SalI + pbs3C5</p><p> |
| - | Pcons+RBS+SalI + pbs3C5 | + | Pcons+RBS+NcoI + pbs3C5</p><p> |
| - | Pcons+RBS+NcoI + pbs3C5 | + | Pcons+RBS+NdeI + pbs3C5</p><p> |
| - | Pcons+RBS+NdeI + pbs3C5 | + | Pcons+RBS+XhoI+RBS+BFP+Ter + pbs1C3</p><p> |
| - | Pcons+RBS+XhoI+RBS+BFP+Ter + pbs1C3 | + | Pcons+RBS+NdeI+RBS+BFP+Ter + pbs1C3</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter + pbs1C3 | + | Pcons+RBS+SacI + pbs1C3</p><p> |
| - | Pcons+RBS+SacI + pbs1C3 | + | Pcons+RBS+NcoI + pbs1C3</p><p> |
| - | Pcons+RBS+NcoI + pbs1C3 | + | |
| - | DNA sequencing of: | + | DNA sequencing of:</p><p> |
| - | SalI-4 | + | SalI-4</p><p> |
| - | HindIII-4 | + | HindIII-4</p><p> |
| - | Pcons+RBS+SalI-3+RBS+BFP+Ter | + | Pcons+RBS+SalI-3+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+SalI-2 | + | Pcons+RBS+SalI-2</p><p> |
| - | Pcons+RBS+NheI-4 | + | Pcons+RBS+NheI-4</p><p> |
| - | NheI(9/26) | + | NheI(9/26)</p><p> |
| - | Pcons+RBS+HindIII+RBS+BFP+Ter(9/30) | + | Pcons+RBS+HindIII+RBS+BFP+Ter(9/30)</p><p> |
| - | Pcons+RBS+SalI+RBS+BFP+Ter(9/30) | + | Pcons+RBS+SalI+RBS+BFP+Ter(9/30)</p> |
| - | + | <p> | |
| - | + | 9/30</p><p> | |
| - | + | PCR and Electrophoresis:</p><p> | |
| - | PCR and Electrophoresis: | + | Pcons+RBS+NdeI+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NdeI+RBS+BFP+Ter | + | NcoI</p><p> |
| - | NcoI | + | Pcons+RBS+NheI+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NheI+RBS+BFP+Ter | + | Pcons+RBS+XhoI+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+XhoI+RBS+BFP+Ter | + | Pcons+RBS+SacI+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+SacI+RBS+BFP+Ter | + | Cultivation of:</p><p> |
| - | Cultivation of: | + | Pcons+RBS+NheI-11+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NheI-11+RBS+BFP+Ter | + | Pcons+RBS+NheI-13+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NheI-13+RBS+BFP+Ter | + | Pcons+RBS+NdeI-3+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NdeI-3+RBS+BFP+Ter | + | Pcons+RBS+NdeI-10+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+NdeI-10+RBS+BFP+Ter | + | Pcons+RBS+XhoI-3+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+XhoI-3+RBS+BFP+Ter | + | Pcons+RBS+XhoI-5+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+XhoI-5+RBS+BFP+Ter | + | Pcons+RBS+SacI-2+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+SacI-2+RBS+BFP+Ter | + | Pcons+RBS+SacI-11+RBS+BFP+Ter</p><p> |
| - | Pcons+RBS+SacI-11+RBS+BFP+Ter | + | NcoI-3</p><p> |
| - | NcoI-3 | + | NcoI-12</p> |
| - | NcoI-12 | + | |
| - | </p> | + | |
<br></div> | <br></div> | ||
Revision as of 14:46, 13 October 2014
- Sunday
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- Saturday
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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
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
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)
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
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
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)
2014/9/16
Transformation of
psb3K3 *2
psb3C3
Digestion of:
psb3K3 dig(EP)
psb1C3 dig(EP)
Cultivation of:
Pcons+RBS+PBAN+RBS+BFP+Ter x8
9/17
Mini prep of:
PBAN：XhoI
SalI
SacI
KpnI
Ndel
AseI
NcoI
NheI
HindIII (fail)
Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4
Ndel-8
HindIII-4 (fail)
SacI-9(fail)
KpnI-1
NcoI-2
NheI-1
Digestion of:
PBAN：XhoI
SalI
SacI
KpnI
Ndel
AseI
NcoI
AseI
Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4
Ndel-8
KpnI-1
NcoI-2
NheI-1
Electrophoresis:
Pcons+RBS+PBAN+RBS+BFP+Ter (Ndel)
Pcons+RBS+PBAN+RBS+BFP+Ter (NcoI)
Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)
Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)
Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)
Pcons+RBS+PBAN+RBS+BFP+Ter (NheI)
Pcons+RBS+PBAN+RBS+BFP+Ter (KpnI)
Pcons+RBS+PBAN+RBS+BFP+Ter (AseI)
PBAN AseI
PBAN Ndel
PBAN KpnI
PBAN SacI
PBAN SalI
PBAN XhoI
Pcons+RBS+XhoI-1 dig(EP)
Pcons+RBS+Xhol-1 dig(ES)
psb1C3 dig(ES)
Cultivation of:
Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)-4.12
Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)-6
Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)-9
Digestion of:
B0034 Kr dig(ES)
Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)
Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)
NcoI Kr dig(EP)
Ndel Kr dig(EP)
SalI Kr dig(EP)
9/18
Electrophoresis:
Well 1：Marker
Well2：B0034 Kr dig(ES)
Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)
Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)
Well5：NcoI Kr dig(EP)
Well6：Ndel Kr dig(EP)
Well7：SalI Kr dig(EP)
Electrophoresis:
Well1：Marker
Well2：B0034 Kr dig(ES)
Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)
Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)
Well5：NcoI Kr dig(EP)
Well6：Ndel Kr dig(EP)
Well7：SalI Kr dig(EP)
Mini prep:
Pcons+RBS+HindIII+RBS+BFP+Ter-6
Pcons+RBS+SalI+RBS+BFP+Ter-9
Pcons+RBS+SacI+RBS+BFP+Ter-4
Pcons+RBS+SacI+RBS+BFP+Ter-11
Ligation of:
Pcons+RBS+AseI+RBS+BFP+Ter Cr
Pcons+RBS+NdeI+RBS+BFP+Ter Cr
NheI Cr
NcoI Cr
9/20
Transformation of:
AseI mini Cr
Electrophoresis:
Well1：Marker
Well2：HindIII-2 Kr dig(EP)
Well3：SacI Kr dig(EP)
Well4：Pcons+RBS+HindIII-6+RBS+BFP+Ter Kr dig(EP)
Well5：Pcons+RBS+SacI-11+RBS+BFP+Ter Kr dig(EP)
Well6：Pcons+RBS+SacI-9+RBS+BFP+Ter Kr dig(EP)
Well7：RBS+BFP+Ter Ar dig(XP)
Well8：Pcons+RBS+NheI+RBS+BFP+Ter Kr dig(EP)
9/21
Electrophoresis of PCR products:
Gel 1
Well1：Marker
Well2：3H1
Well3：3H2
Well4：3H3
Well5：3H4
Well6：3H5
Well7：3H6
Well8：Marker
Well9：6K1
Well10：6K2
Well11：6K3
Well12：6K4
Well13：6K5
Well14：6K6
Well15：3Sl1
Well16：3Sl2
Well17：3Sl3
Gel 2
Well1：Marker
Well2：6Sl4
Well3：6Sl5
Well4：3Nh1
Well5：3Nh2
Well6：3Nh3
Well7：3Nh4
Well8：Marker
Well9：3Nh5
Well10：3Nh6
Well11：6X1
Well12：6X2
Well13：6X3
Well14：6X4
Well15：6X5
Well16：6X6
Well17：1K1
Gel 3
Well1：Marker
Well2：1K2
Well3：1K3
Well4：1K4
Well5：1K5
Well6：1K6
Well7：1X1
Well8：Marker
Well9：1X2
Well10：1X3
Well11：1X4
Well12：1X5
Well13：1X6
Well14：3K1
Well15：3K2
Well16：3K3
Well17：3K4
Gel 4
Well1：Marker
Well2：3K5
Well3：3K6
Well4：3Sc1
Well5：3Sc2
Well6：3Sc3
Well7：3Sc4
Well8：Marker
Well9：3Sc5
Well10：3Sc6
Well11：1Sc1
Well12：1Sc2
Well13：1Sc3
Well14：1Sc4
Well15：1Sc5
Well16：1Sc6
Well17：1Nc1
Gel 5
Well1：Marker
Well2：1Nc2
Well3：1Nc3
Well4：1Nc4
Well5：1Nc5
Well6：1Nc6
Well7：1A1
Well8：Marker
Well9：1A2
Well10：1A3
Well11：1A4
Well12：1A5
Well13：1A6
Well14：1Nd1
Well15：1Nd2
Well16：1Nd3
Well17：1Nd4
Gel 6
Well1：Marker
Well2：1Nd5
Well3：1Nd6
Well4：3X1
Well5：3X2
Well6：3X3
Well7：3X4
Well8：Marker
Well9：3X5
Well10：3X6
Well11：3A1
Well12：3A2
Well13：3A3
Well14：3A4
Well15：3A5
Well16：3A6
Cultivation of:
KpnI-1
NdeI-1
XhoI-1
SacI-1
AseI-1
Pcons+RBS+HindIII-1
Pcons+RBS+KpnI-4
Pcons+RBS+NheI-4
Pcons+RBS+SalI-5
Pcons+RBS+AseI-5
Pcons+RBS+XhoI-1
Pcons+RBS+SacI-1
Pcons+RBS+KpnI-1+RBS+BFP+Ter
Pcons+RBS+XhoI-6+RBS+BFP+Ter
Digestion of:
SalI Kr dig(EP)
Ligation of:
NdeI dig(EP)+C backbone
9/22
Transformation of:
NheI Cr
Pcons+RBS+NheI+RBS+BFP+Ter Cr
Pcons+RBS+NcoI+RBS+BFP+Ter Cr
Pcons+RBS+Ndel+RBS+BFP+Ter Cr
Pcons+RBS+AseI+RBS+BFP+Ter Cr
Digestion of:
HindIII(EP)
Cultivation of:
SacI-3 Kr
NcoI-7 Cr
NcoI-8 Cr
Pcons+RBS+SacI-2
Pcons+RBS+XhoI-5+RBS+BFP+Ter
pbs3C5
Ligation of:
Pcons+RBS+NcoI+RBS+BFP+Ter Cr
Pcons+RBS+HindIII+RBS+BFP+Ter Cr
Pcons+RBS+SacI+RBS+BFP+Ter Cr
Pcons+RBS+SalI+RBS+BFP+Ter Cr
9/23
Electrophoresis:
Well1：Marker
Well2：KpnI-1 Cr dig(EP)
Well3：NdeI-1 Cr dig(EP)
Well4：XhoI-1 Cr dig(EP)
Well5：SacI-1 Cr dig(EP)
Well6：AseI-1 Cr dig(EP)
Well7：Pcons+RBS+HindIII-1 dig(EP)
Well8：Pcons+RBS+KpnI-4 dig(EP)
Well9：Pcons+RBS+NheI-4 dig(EP)
Well10：Pcons+RBS+AseI-5 dig(EP)
Well11：Pcons+RBS+XhoI-1 dig(EP)
Well12：Pcons+RBS+SacI-1 dig(EP)
Well13：Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)
Well14：SalI Kr dig(EP) 9/21
Well15：SalI Kr dig(EP) 9/19
DNA sequencing of:
KpnI-1 Cr dig(EP)
NdeI-1 Cr dig(EP)
XhoI-1 Cr dig(EP)
SacI-1 Cr dig(EP)
AseI-1 Cr dig(EP)
Pcons+RBS+HindIII-1 dig(EP)
Pcons+RBS+KpnI-4 dig(EP)
Pcons+RBS+NheI-4 dig(EP)(fail)
Pcons+RBS+AseI-5 dig(EP)
Pcons+RBS+XhoI-1 dig(EP)
Pcons+RBS+SacI-1 dig(EP)
Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)
Transformation of:
Pcons+RBS+HindIII+RBS+BFP+Ter
Pcons+RBS+NcoI+RBS+BFP+Ter
Pcons+RBS+SalI+RBS+BFP+Ter
Pcons+RBS+SacI+RBS+BFP+Ter
9/24
Cultivation of:
Pcons+RBS+NheI+RBS+BFP+Ter Cr
Pcons+RBS+AseI+RBS+BFP+Ter Cr
Pcons+RBS+NcoI+RBS+BFP+Ter Cr
Pcons+RBS+NdeI+RBS+BFP+Ter Cr
NheI Cr
psb3C5 Cr
Cultivation of:
Pcons+RBS+NheI+RBS+BFP+Ter Cr
Pcons+RBS+AseI+RBS+BFP+Ter Cr
Pcons+RBS+NcoI+RBS+BFP+Ter Cr
Pcons+RBS+NdeI+RBS+BFP+Ter Cr
NheI Cr
psb3C5 Cr
Electrophoresis:
Well1：Marker
Well2：SalI Kr dig(EP)
Well3：Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)
Well4：Pcons+RBS+SacI Cr dig(EP)
Well5：NcoI Cr dig(EP)
Digestion of:
Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)
Pcons+RBS+SacI-2 Cr dig(EP)
NcoI-8 Cr dig(EP)
SalI-3 Kr dig(EP)
Ligation of:
HindIII+psb1C3
PCR of:
SacI x8
SalI x5
HindIII x2
NheI x8
Mini prep and digestion of:
Pcons+RBS+AseI+RBS+BFP+Ter
Pcons+RBS+NheI+RBS+BFP+Ter
Pcons+RBS+NdeI+RBS+BFP+Ter
Pcons+RBS+NcoI+RBS+BFP+Ter
psb3C5
NheI
9/25
Electrophoresis and DNAsequencing:
Well1：Marker
Well2：NheI (fail)
Well3：Pcons+RBS+AseI+RBS+BFP+Ter
Well4：Pcons+RBS+NcoI+RBS+BFP+Ter
Well5：Pcons+RBS+NdeI+RBS+BFP+Ter (fail)
Transformation of:
HindIII
SalI
Pcons+RBS+SacI+RBS+BFP+Ter
Pcons+RBS+SalI+RBS+BFP+Ter
Pcons+RBS+HindIII+RBS+BFP+Ter
PCR of:
NcoI x5(9.10.11.12.13)
Pcons+RBS+SacI x5(7.8.9.10.11)
Cultivation of:
NheI
Pcons+RBS+SacI-2(9/22)
Pcons+RBS+NcoI-8(9/22)
Pcons+RBS+NheI-4
Pcons+RBS+NdeI+RBS+BFP+Ter x2
Pcons+RBS+NheI+RBS+BFP+Ter x2
9/26
Mini prep and digestion(EP) of:
NcoI
NheI
Pcons+RBS+SacI-2
Pcons+RBS+NheI-4
Pcons+RBS+NdeI-1+RBS+BFP+Ter
Pcons+RBS+NdeI-2+RBS+BFP+Ter
Electrophoresis:
Well1：Marker
Well2：Pcons+RBS+NdeI-1+RBS+BFP+Ter
Well3：Pcons+RBS+NdeI-2+RBS+BFP+Ter
Well4：NcoI
Well5：NheI
Well6：Pcons+RBS+NheI-4
Well7：Pcons+RBS+SacI-2
Cultivation of:
psb3C5 x2
HindIII-3
HindIII-4
SalI-3
SalI-4
Pcons+RBS+HindIII-3+RBS+BFP+Ter
Pcons+RBS+HindIII-4+RBS+BFP+Ter
Pcons+RBS+SalI-3+RBS+BFP+Ter
Pcons+RBS+SalI-4+RBS+BFP+Ter
Pcons+RBS+SacI-3+RBS+BFP+Ter
Pcons+RBS+SacI-4+RBS+BFP+Ter
Pcons+RBS+NcoI-3+RBS+BFP+Ter
Pcons+RBS+NcoI-4+RBS+BFP+Ter
9/28
Electrophoresis:
Well1：HindIII-3
Well2：HindIII-4
Well3：SalI-3
Well4：SalI-4
Well5：Pcons+RBS+SacI-3+RBS+BFP+Ter
Well6：Marker
Well7：Pcons+RBS+NcoI-3+RBS+BFP+Ter
Well8：Pcons+RBS+NcoI-4+RBS+BFP+Ter
Well9：Pcons+RBS+SalI-3+RBS+BFP+Ter
Well10：Pcons+RBS+SalI-4+RBS+BFP+Ter
Well11：psb3C5
9/29
Electrophoresis:
Well1：Marker
Well2：Pcons+RBS+HindIII+RBS+BFP+Ter
Well3：Pcons+RBS+HindIII+RBS+BFP+Ter
Well4：Pcons+RBS+HindIII+RBS+BFP+Ter
Well5：Pcons+RBS+HindIII+RBS+BFP+Ter
Well6：Pcons+RBS+HindIII+RBS+BFP+Ter
Well7：Pcons+RBS+HindIII+RBS+BFP+Ter
Well8：Pcons+RBS+NcoI+RBS+BFP+Ter
Well9：Pcons+RBS+NcoI+RBS+BFP+Ter
Well10：Pcons+RBS+NcoI+RBS+BFP+Ter
Well11：Pcons+RBS+NcoI+RBS+BFP+Ter
Well12：Pcons+RBS+NcoI+RBS+BFP+Ter
Well13：Pcons+RBS+NcoI+RBS+BFP+Ter
Well14：Pcons+RBS+SalI+RBS+BFP+Ter
Well15：Pcons+RBS+SalI+RBS+BFP+Ter
Well16：Pcons+RBS+SalI+RBS+BFP+Ter
Well17：Pcons+RBS+SalI+RBS+BFP+Ter
Well18：Pcons+RBS+SalI+RBS+BFP+Ter
Well19：Pcons+RBS+SalI+RBS+BFP+Ter
Well20：psb3C5 dig
Well21：Marker
Ligation of:
Pcons+RBS+KpnI + pbs3C5
Pcons+RBS+SacI + pbs3C5
Pcons+RBS+AseI + pbs3C5
Pcons+RBS+HindIII + pbs3C5
Pcons+RBS+NheI + pbs3C5
Pcons+RBS+SalI + pbs3C5
Pcons+RBS+NcoI + pbs3C5
Pcons+RBS+NdeI + pbs3C5
Pcons+RBS+XhoI+RBS+BFP+Ter + pbs1C3
Pcons+RBS+NdeI+RBS+BFP+Ter + pbs1C3
Pcons+RBS+SacI + pbs1C3
Pcons+RBS+NcoI + pbs1C3
DNA sequencing of:
SalI-4
HindIII-4
Pcons+RBS+SalI-3+RBS+BFP+Ter
Pcons+RBS+SalI-2
Pcons+RBS+NheI-4
NheI(9/26)
Pcons+RBS+HindIII+RBS+BFP+Ter(9/30)
Pcons+RBS+SalI+RBS+BFP+Ter(9/30)
9/30
PCR and Electrophoresis:
Pcons+RBS+NdeI+RBS+BFP+Ter
NcoI
Pcons+RBS+NheI+RBS+BFP+Ter
Pcons+RBS+XhoI+RBS+BFP+Ter
Pcons+RBS+SacI+RBS+BFP+Ter
Cultivation of:
Pcons+RBS+NheI-11+RBS+BFP+Ter
Pcons+RBS+NheI-13+RBS+BFP+Ter
Pcons+RBS+NdeI-3+RBS+BFP+Ter
Pcons+RBS+NdeI-10+RBS+BFP+Ter
Pcons+RBS+XhoI-3+RBS+BFP+Ter
Pcons+RBS+XhoI-5+RBS+BFP+Ter
Pcons+RBS+SacI-2+RBS+BFP+Ter
Pcons+RBS+SacI-11+RBS+BFP+Ter
NcoI-3
NcoI-12
