Team:NCTU Formosa/Notebook/test6

2014/9/4(四) 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(五) 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(二) 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(三) 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(四) Digestion of: Pcons+RBS+PBAN(ES)(8種除了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養的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(五) 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(六) 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)