Contents 1 Labwork overview 1.1 Bba_K1155000 1.2 Bba_K1155001 1.3 Bba_K1155002 1.4 Bba_K1155003 1.5 Bba_K1155004 1.6 Bba_K1155005 1.7 Bba_K1155006 1.8 Bba_K1155007 1.9 Bba_K1155008 1.10 Bba_K1155009 1.11 PSB3K3

Labwork overview

Bba_K1155000

07/01/13

• Digestion of PSB1C3 plasmid and PCR products : Pfnr by EcoRI/ PstI
• Ligation of PSB1C3 and Pfnr

07/02/13

• Tranformation of Bba_K1155000 in DH5α

07/03/13

• Colony PCR of Bba_K1155000 to check good insertion of Pfnr in PSB1C3
• New culture of Bba_K1155000

07/04/13

• Estimation of Pfnr Colony PCR size fragments thanks to Gene manager
• Electrophoresis of Colony PCR products
• Stock of Bba_K1155000
• Culture of Bba_K1155000
• Extraction of Bba_K1155000 from DH5α
• Digestion of Bba_K1155000 by NotI, MluI, HpaI to check good insertion of Pfnr in PSB1C3 again

07/05/13

• Electrophoresis of the digestion of Bba_K1155000 by NotI, HpaI, MluI
• Extraction of Bba_K1155000 from DH5α

07/15/13

• Sequensis analysis of Bba_K1155000

WE OBTAIN OUR BIOBRICK

Bba_K1155001

07/09/13

• PCR purification of BphR1
• Digestion PCR products : BphR1 and digestion of PSB1C3
• Denaturation of EcoRI/PstI used for the digestion of PCR products : BphR1 and PSB1C3
• Ligation of BphR1 and PSB1C3

07/10/13

• Transformation of ligation of PSB1C3 and BphR1

07/11/13

• Colony PCR of Bba_K1155001 in DH5α to check good insertion of BphR1 in PSB1C3
• Electrophoresis of Colony PCR products : Bba_K1155001 in PSB1C3

07/12/13

• Stock of clones 5, 6 for Bba_K1155001
• Extraction of clones 5, 6 for Bba_K1155001 from DH5α

07/15/13

• Digestion of Bba_K1155001 by EcoRI/PstI and SacII to check the size of fragments
• PCR of digestion of Bba_K1155001

07/16/13

• Electrophoresis of the digestion of Bba_K1155001

Bba_K1155002

07/09/13

• PCR purification of BphA1
• Digestion PCR products : BphA1 and digestion of PSB1C3
• Denaturation of EcoRI/PstI used for the digestion of PCR products : BphA1 and PSB1C3
• Ligation of BphA1 and PSB1C3

07/10/13

• Transformation of ligation of PSB1C3 and BphA1

07/11/13

• Colony PCR of Bba_K1155002 in DH5α to check good insertion of BphA1 in PSB1C3
• Electrophoresis of Colony PCR products : Bba_K1155002 in PSB1C3

07/12/13

• Stock of clones 5, 6, 7, 8 for Bba_K1155002
• Streak of Bba_K1155002
• Extraction of clones 5, 6, 7, 8 for Bba_K1155002 from DH5α

07/15/13

• Digestion of Bba_K1155002 by EcoRI/PstI and SacII to check the size of fragments
• PCR of digestion of Bba_K1155002

07/16/13

• Electrophoresis of the digestion of Bba_K1155002

Bba_K1155003

07/02/13

• Tranformation of Bba_K592009 in DH5α

07/03/13

• New culture of Bba_K592009

07/05/13

• Extraction of Bba_K592009 from DH5α

07/08/13

• Digestion of Bba_K592009 by NotI, XhoI, EcoRI, EcoRI/PstI
• Electrophoresis of the digestion of Bba_K592009 by NotI, XhoI, EcoRI, EcoRI/PstI

07/12/13

• Transformation of Bba_B0015, Bba_B0017
• Design of RBS-AmilCP oligos

07/15/13

• Liquid culture of Bba_B0015, Bba_B0017

07/16/13

• Extraction of Bba_B0015, Bba_B0017 from DH5α
• Digestion of Bba_B0015, Bba_B0017, Bba_I732019 by EcoRI/PstI

Bba_K1155004

Bba_K1155005

Bba_K1155006

Bba_K1155007

07/02/13

• Tranformation of Bba_I732017 in DH5α

07/03/13

• New culture of Bba_I732017

07/05/13

• Extraction of Bba_I732017 from DH5α

07/08/13

• Digestion of Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI
• Electrophoresis of the digestion of Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI

07/10/13

• Transformation of Bba_I732019 in DH5α

07/12/13

• Transformation of Bba_I732019, Bba_B0015, Bba_B0017

07/15/13

• Liquid culture of Bba_B0015, Bba_B0017, Bba_I732019

07/16/13

• Extraction of Bba_B0015, Bba_B0017, Bba_I732019 from DH5α
• Digestion of Bba_B0015, Bba_B0017, Bba_I732019 by EcoRI/PstI

Bba_K1155008

Bba_K1155009

07/01/13

• Design of oligos for amplification of BphR2 gene of Pseudomonas pseudoalcaligenes, Pseudomonas oleovorans

07/09/13

• PCR purification of BphR2
• Digestion PCR products : BphR2 and digestion of PSB1C3
• Denaturation of EcoRI/PstI used for the digestion of PCR products : BphR2 and PSB1C3
• Ligation of BphR2 and PSB1C3

07/10/13

• Transformation of ligation of PSB1C3 and BphR2

07/11/13

• Colony PCR of BphR2 in PSB1C3 in DH5α to check good insertion of BphR2 in PSB1C3
• Electrophoresis of Colony PCR products : BphR2 in PSB1C3

07/12/13

• Stock of clones 3, 4 for BphR2
• Streak of BphR2
• Extraction of clones 3, 4 for BphR2 from DH5α

07/15/13

• Digestion of BphR2 in PSB1C3 by EcoRI/PstI and SacII to check the size of fragments
• PCR of digestion of BphR2 in PSB1C3

07/16/13

• Electrophoresis of the digestion of BphR2 in PSB1C3

PSB3K3

07/09/13

• Transformation of Bba_J04450 in DH5α

07/10/13

• Mini and maxi preparation of the culture of Bba_J04450

07/11/13

• Extraction of Bba_J04450 from DH5α
• Digestion of Bba_J04450 to chek the size for the plasmid
• Electrophoresis of the digestion of Bba_J04450

07/16/13

• Digestion of PSB3K3 by EcoRI/PstI
• Electrophoresis of the digestion of PSB3K3 by EcoRI/PstI
• Gel purification of electrophoresis of the digestion of PSB3K3 by EcoRI/PstI