Team:NCTU Formosa/Notebook/test6

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)

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

2014/9/17 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)

2014/9/18 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) 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

2014/9/20 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)