"
Team:Paris Saclay/Labwork
From 2013.igem.org
(Difference between revisions)
(→BBa_K1155007) |
(→BBa_K1155007) |
||
| (8 intermediate revisions not shown) | |||
| Line 195: | Line 195: | ||
08/07/13 | 08/07/13 | ||
* Sequensis analysis of BBa_K1155003 [File:Psright.png|20px]] | * Sequensis analysis of BBa_K1155003 [File:Psright.png|20px]] | ||
| + | |||
| + | '''WE OBTAIN OUR BIOBRICK.''' | ||
=='''BBa_K1155004'''== | =='''BBa_K1155004'''== | ||
| Line 364: | Line 366: | ||
07/30/13 | 07/30/13 | ||
* Digestion of BBa_I732017 by EcoRI/PstI | * Digestion of BBa_I732017 by EcoRI/PstI | ||
| - | + | * Digestion of BBa_I732017 by EcoRI/SpeI | |
| - | + | * Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI | |
| - | * Digestion of BBa_I732017 by EcoRI/ | + | * Gel purification of digestion of BBa_I732017 by EcoRI/SpeI [[File:Pswrong.png|20px]] |
| - | * Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI | + | |
| - | * Gel purification of digestion of BBa_I732017 by EcoRI/ | + | |
07/31/13 | 07/31/13 | ||
| Line 412: | Line 412: | ||
08/01/13 | 08/01/13 | ||
| - | * Gel purification of PCR products : FNR Part I, FNR Part II, RBS-FNR Part I | + | * Gel purification of PCR products : FNR Part I, FNR Part II, RBS-FNR Part I pSB1C3 [[File:Psright.png|20px]] |
* Gibson assembly | * Gibson assembly | ||
| Line 445: | Line 445: | ||
08/27/13 | 08/27/13 | ||
* Electrophoresis of FNR Part I , FNR Part II, RBS-FNR Part I [[File:Psright.png|20px]] | * Electrophoresis of FNR Part I , FNR Part II, RBS-FNR Part I [[File:Psright.png|20px]] | ||
| - | * Electrophoresis of | + | * Electrophoresis of pSB1C3 [[File:Pswrong.png|20px]] |
| - | * Digestion of | + | * Digestion of pBB1C3 by DnpI to clean it |
| - | * Electrophoresis of the digestion of | + | * Electrophoresis of the digestion of pSB1C3 by DnpI [[File:Psright.png|20px]] |
08/28/13 | 08/28/13 | ||
| - | * Gel purification of | + | * Gel purification of pSB1C3 digested by DnpI |
| - | * Electrophoresis to check the gel purification of | + | * Electrophoresis to check the gel purification of pSB1C3 digested by DnpI [[File:Psright.png|20px]] |
* Gibson assembly of RSB-FRN, FNR, | * Gibson assembly of RSB-FRN, FNR, | ||
* Transformation of FNR, RBS-FNR in DH5α [[File:Psright.png|20px]] | * Transformation of FNR, RBS-FNR in DH5α [[File:Psright.png|20px]] | ||
| Line 468: | Line 468: | ||
07/09/13 | 07/09/13 | ||
* PCR purification of BphR2 | * PCR purification of BphR2 | ||
| - | * Digestion PCR products : BphR2 and digestion of | + | * Digestion PCR products : BphR2 and digestion of pSB1C3 |
| - | * Denaturation of EcoRI/PstI used for the digestion of PCR products : BphR2 and | + | * Denaturation of EcoRI/PstI used for the digestion of PCR products : BphR2 and pSB1C3 |
| - | * Ligation of BphR2 and | + | * Ligation of BphR2 and pSB1C3 |
07/10/13 | 07/10/13 | ||
| - | * Transformation of ligation of | + | * Transformation of ligation of pSB1C3 and BphR2 [[File:Psright.png|20px]] |
07/11/13 | 07/11/13 | ||
| - | * Colony PCR of BphR2 in | + | * Colony PCR of BphR2 in pSB1C3 in DH5α to check good insertion of BphR2 in pSB1C3 |
| - | * Electrophoresis of Colony PCR products : BphR2 in | + | * Electrophoresis of Colony PCR products : BphR2 in pSB1C3 [[File:Psright.png|20px]] |
07/12/13 | 07/12/13 | ||
| Line 485: | Line 485: | ||
07/15/13 | 07/15/13 | ||
| - | * Digestion of BphR2 in | + | * Digestion of BphR2 in pSB1C3 by EcoRI/PstI and SacII to check the size of fragments |
| - | * PCR of digestion of BphR2 in | + | * PCR of digestion of BphR2 in pSB1C3 |
07/16/13 | 07/16/13 | ||
| - | * Electrophoresis of the digestion of BphR2 in | + | * Electrophoresis of the digestion of BphR2 in pSB1C3 [[File:Pswrong.png|20px]] |
07/30/13 | 07/30/13 | ||
| Line 499: | Line 499: | ||
08/01/13 | 08/01/13 | ||
| - | * Gel purification of PCR products : BphR2 Part I, BphR2 Part II | + | * Gel purification of PCR products : BphR2 Part I, BphR2 Part II, pSB1C3 [[File:Psright.png|20px]] |
* Gibson assembly | * Gibson assembly | ||
* PCR of : RBS-BphR2 Part I | * PCR of : RBS-BphR2 Part I | ||
| Line 552: | Line 552: | ||
08/27/13 | 08/27/13 | ||
* Electrophoresis of RBS-BphR2 Part I, BphR2 Part II [[File:Psright.png|20px]] | * Electrophoresis of RBS-BphR2 Part I, BphR2 Part II [[File:Psright.png|20px]] | ||
| - | * Electrophoresis of | + | * Electrophoresis of pSB1C3 [[File:Pswrong.png|20px]] |
| - | * Digestion of | + | * Digestion of pSB1C3 by DnpI tto clean it |
| - | * Electrophoresis of the digestion of | + | * Electrophoresis of the digestion of pSB1C3 by DnpI [[File:Psright.png|20px]] |
08/28/13 | 08/28/13 | ||
| - | * Gel purification of | + | * Gel purification of pSB1C3 digested by DnpI |
| - | * Electrophoresis to check the gel purification of | + | * Electrophoresis to check the gel purification of pSB1C3 digested by DnpI [[File:Psright.png|20px]] |
* Gibson assembly of RBS-BphR2 | * Gibson assembly of RBS-BphR2 | ||
* Transformation of RBS-BphR2 in DH5α [[File:Psright.png|20px]] | * Transformation of RBS-BphR2 in DH5α [[File:Psright.png|20px]] | ||
| Line 568: | Line 568: | ||
* Electrophoresis of Colony PCR of RBS-BphR2 [[File:Psright.png|20px]] | * Electrophoresis of Colony PCR of RBS-BphR2 [[File:Psright.png|20px]] | ||
| - | ==''' | + | =='''pSB3K3'''== |
07/09/13 | 07/09/13 | ||
| Line 582: | Line 582: | ||
07/16/13 | 07/16/13 | ||
| - | * Digestion of | + | * Digestion of pSB3K3 by EcoRI/PstI |
| - | * Electrophoresis of the digestion of | + | * Electrophoresis of the digestion of pSB3K3 by EcoRI/PstI [[File:Psright.png|20px]] |
| - | * Gel purification of electrophoresis of the digestion of | + | * Gel purification of electrophoresis of the digestion of pSB3K3 by EcoRI/PstI [[File:Psright.png|20px]] |
08/06/13 | 08/06/13 | ||
| Line 593: | Line 593: | ||
* Electrophoresis of BBa_J004450 digested by EcoRI/Pst1 to check if the digestion [[File:Psright.png|20px]] | * Electrophoresis of BBa_J004450 digested by EcoRI/Pst1 to check if the digestion [[File:Psright.png|20px]] | ||
* Electrophoresis of BBa_J004450 digested by EcoRI/PstI [[File:Psright.png|20px]] | * Electrophoresis of BBa_J004450 digested by EcoRI/PstI [[File:Psright.png|20px]] | ||
| - | * Electroelution of | + | * Electroelution of pSB3K3 digested by EcoRI/PstI [[File:Pswrong.png|20px]] |
08/13/13 | 08/13/13 | ||
| Line 663: | Line 663: | ||
* Digestion of BBa_K1155000 by SpeI/PstI | * Digestion of BBa_K1155000 by SpeI/PstI | ||
* Denaturation of SpeI/PstI used for the digestion of BBa_K1155000 [[File:Psright.png|20px]] | * Denaturation of SpeI/PstI used for the digestion of BBa_K1155000 [[File:Psright.png|20px]] | ||
| - | * Ligation of | + | * Ligation of Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 |
| - | * Transformation of ligation of | + | * Transformation of ligation of Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 [[File:Psright.png|20px]] |
* Purification colony from strain MG1655Z1 Δfnr::Km containing plasmid pcp20 [[File:Psright.png|20px]] | * Purification colony from strain MG1655Z1 Δfnr::Km containing plasmid pcp20 [[File:Psright.png|20px]] | ||
08/27/13 | 08/27/13 | ||
| - | * Colony PCR of ligation | + | * Colony PCR of ligation Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 |
| - | * Electrophoresis of the colony PCR products : | + | * Electrophoresis of the colony PCR products : Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 [[File:Psright.png|20px]] |
| - | * Streak of | + | * Streak of Pndh* with RBS-LacZ-Term in pSB1C3 and Pfnr with RBS-Amil CP-Term in pSB1C3 to purify them [[File:Psright.png|20px]] |
* Culture of strain MG1655Z1 Δfnr::Km containing plasmid pcp20 [[File:Psright.png|20px]] | * Culture of strain MG1655Z1 Δfnr::Km containing plasmid pcp20 [[File:Psright.png|20px]] | ||
08/28/13 | 08/28/13 | ||
| - | * Purification of colony transformed with ligation : | + | * Purification of colony transformed with ligation : Pndh* with RBS-LacZ-Term in pSB1C3, Pndh* with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions [[File:Pswrong.png|20px]] |
* Culture of mutant strain MG1655Z1 Δfnr with LB [[File:Psright.png|20px]] | * Culture of mutant strain MG1655Z1 Δfnr with LB [[File:Psright.png|20px]] | ||
08/29/13 | 08/29/13 | ||
* Purification colony of strain MG1655Z1 Δfnr with kanamycin, chloramphenicol, ampicilin or LB | * Purification colony of strain MG1655Z1 Δfnr with kanamycin, chloramphenicol, ampicilin or LB | ||
| - | * Purification of colony transformed with ligation : | + | * Purification of colony transformed with ligation : Pndh* with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions [[File:Pswrong.png|20px]] |
| - | * Colony PCR of | + | * Colony PCR of Pndh* wit RBS_AmilCP-Term in DH5α |
* Digestion of BBa_K1155000, BBa_K1155003, BBa_K1155007 by Xbal/PstI | * Digestion of BBa_K1155000, BBa_K1155003, BBa_K1155007 by Xbal/PstI | ||
* Electrophoresis to check the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI [[File:Psright.png|20px]] | * Electrophoresis to check the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI [[File:Psright.png|20px]] | ||
* Gel purification of the digestion of Bba_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI | * Gel purification of the digestion of Bba_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI | ||
* Electrophoresis to check the gel purification of the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI [[File:Psright.png|20px]] | * Electrophoresis to check the gel purification of the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI [[File:Psright.png|20px]] | ||
| - | * Liquid culture in aerobic or anaerobic conditions of | + | * Liquid culture in aerobic or anaerobic conditions of Pndh* with RBS-Amil CP-Term in pSB1C3 [[File:Psright.png|20px]] |
'''WE CHARACTERIZE BBa_K1155000. BBa_K1155003 WORKS TOO.''' | '''WE CHARACTERIZE BBa_K1155000. BBa_K1155003 WORKS TOO.''' | ||
| Line 692: | Line 692: | ||
08/30/13 | 08/30/13 | ||
| - | * Ligation of | + | * Ligation of Pndh* with RBS-Amil CP-Term in pSB1C3 |
| - | * Transformation of ligation of | + | * Transformation of ligation of Pndh* with RBS-Amil CP-Term in pSB1C3 in DH5α [[File:Psright.png|20px]] |
| - | * Ligation of | + | * Ligation of Pndh* with RBS-LacZ-Term in pSB3K3 |
| - | * Transformation of ligation of | + | * Transformation of ligation of Pndh* with RBS-LacZ-Term in pSB3K3 and in DH5α [[File:Psright.png|20px]] |
| - | * Transformation of ligation of | + | * Transformation of ligation of Pndh* with RBS-LacZ-Term in pSB3C3 and in DH5α [[File:Pswrong.png|20px]] |
| - | * Electrophoresis of Colony PCR of | + | * Electrophoresis of Colony PCR of Pndh* with RBS_AmilCP-Term in DH5α [[File:Psright.png|20px]] |
* Result of the purification colony of MG1655Z1 Δfnr in ampicilin, chloramphenicol, kanamycin or LB medium [File:Psright.png|20px]] | * Result of the purification colony of MG1655Z1 Δfnr in ampicilin, chloramphenicol, kanamycin or LB medium [File:Psright.png|20px]] | ||
| Line 731: | Line 731: | ||
08/19/13 | 08/19/13 | ||
* EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | * EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | ||
| - | * Ligation of | + | * Ligation of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 |
| - | * Transformation of | + | * Transformation of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α [[File:Psright.png|20px]] |
08/21/13 | 08/21/13 | ||
| - | * Colony PCR of | + | * Colony PCR of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α |
08/26/13 | 08/26/13 | ||
* Digestion of BBa_K1155004 by SpeI/PstI | * Digestion of BBa_K1155004 by SpeI/PstI | ||
* Denaturation of SpeI/PstI used for the digestion of BBa_K1155004 [[File:Psright.png|20px]] | * Denaturation of SpeI/PstI used for the digestion of BBa_K1155004 [[File:Psright.png|20px]] | ||
| - | * Ligation of | + | * Ligation of PnirB clone 6 with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 |
| - | * Transformation of ligation of | + | * Transformation of ligation of PnirB with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 [[File:Psright.png|20px]] |
08/27/13 | 08/27/13 | ||
| - | * Colony PCR of ligation | + | * Colony PCR of ligation PnirB with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3 |
| - | * Electrophoresis of the colony PCR products : | + | * Electrophoresis of the colony PCR products : PnirB with RBS-LacZ-Term [[File:Pswrong.png|20px]] |
| - | * Electrophoresis of the colony PCR products : | + | * Electrophoresis of the colony PCR products : PnirB with RBS-Amil CP-Term in pSB1C3 [[File:Psright.png|20px]] |
| - | * Streak of | + | * Streak of PnirB with RBS-Amil CP-Term in pSB1C3 to purify them [[File:Psright.png|20px]] |
08/28/13 | 08/28/13 | ||
| - | * Purification of colony transformed with ligation : | + | * Purification of colony transformed with ligation : PnirB with RBS-LacZ-Term in pSB1C3, PnirB with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions [[File:Pswrong.png|20px]] |
08/29/13 | 08/29/13 | ||
| - | * Purification of colony transformed with ligation : | + | * Purification of colony transformed with ligation : PnirB with RBS-LacZ-Term in pSB1C3 by streaking in aerobic or anaerobic conditions [[File:Pswrong.png|20px]] |
* Electrophoresis to check the digestion of BBa_K1155004 by SpeI/PstI and plasmids already digested by SpeI and after digested by PstI [[File:Pswrong.png|20px]] | * Electrophoresis to check the digestion of BBa_K1155004 by SpeI/PstI and plasmids already digested by SpeI and after digested by PstI [[File:Pswrong.png|20px]] | ||
| Line 782: | Line 782: | ||
08/19/13 | 08/19/13 | ||
* EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | * EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | ||
| - | * Ligation of | + | * Ligation of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 |
| - | * Transformation of | + | * Transformation of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α [[File:Psright.png|20px]] |
08/21/13 | 08/21/13 | ||
| - | * Colony PCR of | + | * Colony PCR of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α |
08/26/13 | 08/26/13 | ||
| Line 820: | Line 820: | ||
08/19/13 | 08/19/13 | ||
* EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | * EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term | ||
| - | * Ligation of | + | * Ligation of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 |
| - | * Transformation of | + | * Transformation of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α [[File:Psright.png|20px]] |
08/21/13 | 08/21/13 | ||
| - | * Colony PCR of | + | * Colony PCR of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α |
08/26/13 | 08/26/13 | ||
| Line 838: | Line 838: | ||
08/30/13 | 08/30/13 | ||
| - | * Ligation of | + | * Ligation of PnarK with RBS-LacZ-Term in pSB3K3 |
| - | * Transformation of ligation of | + | * Transformation of ligation of PnarK with RBS-LacZ-Term in pSB3K3 in DH5α [[File:Psright.png|20px]] |
'''BBa_K1155006 WORKS IN ANAEROBIC CONDITIONS. BBa_K1155007 WORKS.''' | '''BBa_K1155006 WORKS IN ANAEROBIC CONDITIONS. BBa_K1155007 WORKS.''' | ||
Latest revision as of 00:59, 5 October 2013
Labwork overview
BBa_K1155000
Engineered promoter region of E. coli Pndh gene that allows repression of the expression of the downstream gene in anaerobic conditions
07/01/13
- PCR of pSB1C3 and Pndh*
- Digestion of pSB1C3 plasmid and Pndh* PCR products by EcoRI/ PstI
- Ligation of pSB1C3 and Pndh*
07/02/13
- Transformation of E. coli DH5α with the ligation mix
07/03/13
- PCR on colonies obtained after the transformation to check good insertion of Pndh* in pSB1C3
- Colonies assayed by PCR streaked on plates and used for liquid cultures
07/04/13
- Estimation of the expected PCR size fragments using bio-informatic tools
- Gel Electrophoresis of PCR products obtained the day before
- Glycerol Stocks of the clones obtained
- DNA Extraction of the plasmids from DH5α
- Digestion of the plasmids by NotI, MluI, HpaI to check with a different method the good insertion of Pndh* in pSB1C3
- New liquid culture of the correct clone
07/05/13
- Electrophoresis of the digestion of the correct clone, now named BBa_K1155000 by NotI, HpaI, MluI
- Extraction of plasmids DNA of BBa_K1155000 from DH5α for sequencing
07/15/13
- Sequence analysis of BBa_K1155000
WE OBTAINED OUR BIOBRICK.
BBa_K1155001
07/09/13
- PCR and purification of PbphbR1 DNA fragment
- Digestion PCR products : PbphR1 and pSB1C3 by EcoRI/PstI
- Denaturation of EcoRI/PstI enzymes used for the digestion of PCR products : PbphR1 and pSB1C3
- Ligation of PbphR1 and pSB1C3 fragments
07/10/13
- Transformation of E. coli with ligation mix (pSB1C3 and PbphR1)
07/11/13
- PCR on colonies obtained after the transformation to check good insertion of PbphR1 in pSB1C3
- Gel electrophoresis of PCR products : clones 5 and 6 correct
07/12/13
- Glycerol stock of clones 5, 6
- DNA plasmid extraction of clones 5, 6
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
- Gel electrophoresis of the digestion of BBa_K1155001
07/17/13
- Glycerol Stock of BBa_K1155001
- DNA plasmid extraction of BBa_K1155001 from DH5α
07/24/13
- Sequence analysis of BBa_K1155001
WE OBTAINED OUR BIOBRICK.
BBa_K1155002
07/09/13
- PCR and purification of PbphA1
- Digestion PCR products : PbphA1 and pSB1C3 by EcoRI/PstI
- Denaturation of EcoRI/PstI enzymes used for the digestion of PCR products : PbphA1 and pSB1C3
- Ligation of PbphA1 and pSB1C3
07/10/13
- Transformation of E. coli DH5α with the ligation mix
07/11/13
- PCR on colonies obtained after the transformation to check good insertion of PbphA1 in pSB1C3
- Gel electrophoresis of PCR products : BBa_K1155002 in PSB1C3
07/12/13
- Stock of clones 5, 6, 7, 8 for BBa_K1155002
- Streak of BBa_K1155002
- DNA plasmid 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
- Gel electrophoresis of the digestion of BBa_K1155002
07/25/13
- New ligation of PbphA1 in pSB1C3
07/26/13
- Transformation of E. coli DH5α with the ligation mix
07/30/13
- PCR on colonies obtained after the transformation to check good insertion of PbphA1 in pSB1C3
- Gel electrophoresis of the PCR products
08/06/13
- Sequence analysis for BBa_K1155002 in clones 4, 17 and 22
WE OBTAIN OUR BIOBRICK.
BBa_K1155003
07/02/13
- Tranformation of E. coli DH5α with BBa_K592009
07/03/13
- New culture of BBa_K592009
07/05/13
- Extraction of BBa_K592009 from E. coli 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 E. coli DH5α with BBa_B0015, BBa_B0017
- Design of RBS-AmilCP primers
07/15/13
- Liquid culture of BBa_B0015, BBa_B0017
07/16/13
- Plasmidic DNA extraction of BBa_B0015, BBa_B0017 from E. coli DH5α
- Digestion of BBa_B0015, BBa_B0017, BBa_I732019 by EcoRI/PstI
07/18/13
- PCR of BBa_B0015, BBa_B0017 digested by EcoRI/PstI
07/19/13
- Gel electrophoresis of PCR products : BBa_B0015, BBa_B0017
- Gel purification of the electrophoresis of PCR products : BBa_B0015, BBa_B0017
07/22/13
- Digestion of BBa_K592009 by SpeI/PstI, BBa_B0015 and BBa_B0017 by PstI/XbaI
- Denaturation of SpeI,PstI and PstI,XbaI enzymes used for the digestion of BBa_K592009, BBa_B0015 and BBa_B0017
07/23/13
- Gel electrophoresis of the digestion of BBa_K592009 by SpeI/PstI
- Gel electrophoresis of the digestion of BBa_B0015, BBa_B0017 by XbaI/PstI
- Digestion of BBa_K592009 by SpeI/PstI
- Gel electrophoresis to check the digestion of BBa_K592009 by SpeI/PstI
- Liquid culture of BBa_K592009 in E. coli DH5α
07/24/13
- Plasmid extraction of BBa_K592009 from E. coli DH5α
07/25/13
- Gel purification of the terminator
- Gel electrophoresis to check the gel purification of Term
- PCR of BBa_K592209 with the primer : RBS-Amil CP and suffix_pSB1C3
- Digestion of PCR products : RBS-Amil CP by EcoRI/SpeI
07/26/13
- Denaturation of EcoRI/SpeI enzymes used for the digestion of PCR product : RBS-AmilCP
- Gel Electrophoresis of the digestion of PCR products : RBS-AmilCP
- Ligation of RBS-AmilCP and Term-pSB1C3
07/30/13
- Tranformation of E. coli DH5α with the ligation mix
08/02/13
- DNA plasmid extraction
- Electrophoresis to check the plasmid extraction
08/05/13
- Digestion of BBa_K1155003 by EcoRI/PstI to check the cloning between RBS-Amil CP,Term and pSB1C3
- Gel electrophoresis of the digestion of BBa_K1155003
08/07/13
- Sequensis analysis of BBa_K1155003 [File:Psright.png|20px]]
WE OBTAIN OUR BIOBRICK.
BBa_K1155004
07/25/13
- PCR to amplify the promoter of nirB
- Gel Electrophoresis to check the correct amplification of PnirB
- PCR to amplify the promoter of nirB
07/26/13
- Gel Electrophoresis to check the correct amplification of PnirB
- PCR of PnirB
- Electrophoresis of PCR products : PnirB
- Gel purification of PCR products : PnirB
- Digestion of PCR products : PnirB by EcoRI/PstI
07/30/13
- Denaturation of EcoRI/SpeI used of the digestion of PCR products : PnirB
07/31/13
- Digestion of PSB1C3 by EcoRI/PstI'
- Denaturation of EcoRI/PstI used for the digestion of PSB1C3
- Ligation of PnirB in pSB1C3
- Electrophoresis to check the ligation of PnirB in pSB1C3
- Transformation of BBa_K1155004
08/05/13
- Colony PCR of BBa_K1155004 in DH5α
08/06/13
- Electrophoresis to check the Colony PCR products : BBa_K1155004
- Liquid culture of DH5α with BBa_K1155004
08/20/13
- Sequences analysis
WE OBTAIN OUR BIOBRICK
BBa_K1155005
07/25/13
- Colony PCR of PnarG
- Electrophoresis to check the colony PCR of PnarG
- Colony PCR of PnarG
07/26/13
- Electrophoresis of PCR products : PnarG
- PCR of PnarG
- Electrophoresis of PCR products : PnarG
- Gel purification of PCR products : PnarG
- Digestion of PCR products : PnarG by EcoRI/PstI
07/30/13
- Denaturation of EcoRI/SpeI used of the digestion of PCR products : PnarG
07/31/13
- Digestion of pSB1C3 by EcoRI/PstI'
- Denaturation of EcoRI/PstI used for the digestion of pSB1C3
- Ligation of PnarG in pSB1C3
- Electrophoresis to check the ligation of PnarG in pSB1C3
- Transformation of BBa_K1155005
08/05/13
- Colony PCR of BBa_K115005 in DH5α
08/06/13
- Electrophoresis to check the Colony PCR products : BBa_K1155005
- Liquid culture of DH5α with BBa_K1155005
08/20/13
- Sequences analysis
WE OBTAIN OUR BIOBRICK
BBa_K1155006
07/25/13
- Colony PCR of PnarK
- Electrophoresis to check the colony PCR of PnarK
- Colony PCR of PnarK
07/26/13
- Electrophoresis of PCR products : PnarK
- PCR of PnarK
- Electrophoresis of PCR products : PnarK
- Gel purification of PCR products : PnarK
- Digestion of PCR products : PnarK by EcoRI/PstI
07/30/13
- Denaturation of EcoRI/SpeI used of the digestion of PCR products : PnarK
07/31/13
- Digestion of pSB1C3 by EcoRI/PstI'
- Denaturation of EcoRI/PstI used for the digestion of pSB1C3
- Ligation of PnarK in pSB1C3
- Electrophoresis to check the ligation of PnarK in pSB1C3
- Transformation of BBa_K1155006
08/05/13
- Colony PCR of BBa_K1155006 in DH5α
08/06/13
- Electrophoresis to check the Colony PCR products : BBa_K1155006
- Liquid culture of DH5α with BBa_K1155006
08/20/13
- Sequences analysis
WE OBTAIN OUR BIOBRICK
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
07/17/13
- Electrophoresis of digestions of BBa_B0015, BBa_B0017 by EcoRI/PstI
- Electrophoresis of digestions of BBa_I732019 by EcoRI/PstI
07/18/13
- PCR of BBa_B0015, BBa_B0017 digested by EcoRI/PstI
07/19/13
- Electrophoresis of Colony PCR products : BBa_B0015, BBa_B0017
- Gel purification of the electrophoresis of Colony PCR products : BBa_B0015, BBa_B0017
07/22/13
- Digestion of BBa_I732017 by EcoRI/SpeI, BBa_B0015 and BBa_B0017 by PstI/XbaI
- Denaturation of EcoRI/SpeI and PstI/XbaI used for the digestion of BBa_I732017, BBa_K502009, BBa_B0015 and BBa_B0017
07/23/13
- Electrophoresis of the digestion of BBa_I732017 by EcoRI/SpeI, BBa_B0015, BBa_B0017 by XbaI/PstI
- Gel purification of digestion products : RBS-LacZ
- Electrophoresis to check the gel purification of digestion products : RBS-LacZ
- Digestion of BBa_I732017 by EcoRI/Pst
- Electrophoresis to check the gel digestion of BBa_I732017 by EcoRI/SpeI
- Liquid culture of BBa_I732017 in DH5α
07/24/13
- Extraction of BBa_I732017 from DH5α
07/25/13
- Gel purification of Term
- Electrophoresis to check the gel purification of Term
07/30/13
- Digestion of BBa_I732017 by EcoRI/PstI
- Digestion of BBa_I732017 by EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI
- Gel purification of digestion of BBa_I732017 by EcoRI/SpeI
07/31/13
- Digestion of BBa_I732017 by EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_I732017 by EcoRI/SpeI
- Gel purification of digestion of RBS-LacZ
08/02/13
- Digestion of BBa_I732017 by EcoRI/SpeI to check sizes of fragments
- Electrophoresis of the digestion of BBa_I732017 by EcoRI/SpeI
08/05/13
- Digestion of BBa_I732017 by EcoRI/SpeI
- Electrophoresis of the digestion of BBa_I732017 by EcoRI/SpeI
- PCR of pSB1C3
- Electrophoresis of PCR of pSB1C3
- Gel purification of electrophoresis of PCR of pSB1C3
08/06/13
- Electroelution of BBa_I732017 digested by EcoRI/SpeI
08/07/13
- Ligation of RBS-LacZ-Term in pSB1C3
- Tranformation BBa_K115507 in DH5α
08/08/13
- Colony PCR of BBa_K115007 in DH5α
- Electrophoresis to check the colony PCR products : Bba_K1155007
08/09/13
- Extraction of BBa_K115007 from DH5α
08/19/13
- Sequence analysis of BBa_K1155007
WE OBTAIN OUR BIOBRICK.
BBa_K1155008
07/30/13
- PCR of : FNR Part I, FNR Part II, RBS-FNR Part I
- Electrophoresis of the PCR products : FNR Part I, FNR Part II, RBS-FNR Part I
08/01/13
- Gel purification of PCR products : FNR Part I, FNR Part II, RBS-FNR Part I pSB1C3
- Gibson assembly
08/02/13
- Tranformation of FNR and RBS-FNR in DH5α
08/08/13
- Extraction of plasmid of FNR, RBS-FNR
08/09/13
- Electrophoresis of the PCR of FNR Part I, FNR Part II, RBS_FNR Part I to check the gel purification
- PCR of FNR Part I, FNR Part II, RBS-FNR Part I
08/12/13
- Electrophoresis of PCR products : RBS-FNR Part I, FNR Part I and FNR Part II
08/13/13
- PCR of FRN Part I, FNR Part II
- Electrophoresis of PCR products : FRN Part I, FNR Part II File:Psright.png|20px]]
- Gel purification of PCR product : FNR Part I, FNR Part II, RBS-FNR Part I
08/14/13
- Gibson assembly of RBS-FNR, FNR
08/21/13
- Transformation of FNR, RBS-FNR in DH5α
08/26/13
- Gibson assembly RBS-FNR, FNR
- Transformation of FNR, RBS-FNR in DH5α
08/27/13
- Electrophoresis of FNR Part I , FNR Part II, RBS-FNR Part I
- Electrophoresis of pSB1C3
- Digestion of pBB1C3 by DnpI to clean it
- Electrophoresis of the digestion of pSB1C3 by DnpI
08/28/13
- Gel purification of pSB1C3 digested by DnpI
- Electrophoresis to check the gel purification of pSB1C3 digested by DnpI
- Gibson assembly of RSB-FRN, FNR,
- Transformation of FNR, RBS-FNR in DH5α
08/29/13
- Colony PCR of FNR, RBS-FNR and RBS-BphR2 in DH5α
08/30/13
- Electrophoresis of Colony PCR of FNR, RBS-FNR
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
07/30/13
- PCR of : BphR2 Part I, BphR2 Part II, RBS-BphR2 Part I
- Electrophoresis of the PCR products : BphR2 Part I, BphR2 Part II
- Electrophoresis of the PCR products : RBS-BphR2 Part I
- PCR of : RBS-BphR2 Part I
- Gel purification of PCR of : RBS-BphR2 Part I
08/01/13
- Gel purification of PCR products : BphR2 Part I, BphR2 Part II, pSB1C3
- Gibson assembly
- PCR of : RBS-BphR2 Part I
- Electrophoresis of the PCR products : RBS-BphR2 Part I
08/02/13
- Tranformation of BphR2 in DH5α
08/08/13
- Extraction of plasmid of BphR2
- Gel purification of RBS-BphR2 Part I [File:Pswrong.png|20px]]
08/09/13
- Electrophoresis of the PCR of BphR2 Part I, BphR2 Part II, RBS_BphR2 Part I to check the gel purification
- PCR of BphR2 Part I, BphR2 Part II, RBS-BphR2 Part I
08/12/13
- Electrophoresis of PCR products : RBS-BphR2 Part I, BphR2 Part I, BphR2 Part II
08/13/13
- PCR of BphR2 Part I
- Electrophoresis of PCR products : BphR2 Part I
- Electrophoresis of PCR product : BphR2 Part I to separate BphR2 from an other stripe
- PCR of BphR2 Part I
- Gel purification of PCR product : BphR2 Part II, RBS-BphR2 Part I
08/14/13
- Gibson assembly of RBS-BphR2
- Electrophoresis of PCR products : BphR2 Part I
- PCR of BphR2 Part I
08/19/13
- Electrophoresis gel of PCR products : BphR2 Part I
- Gel purification of PCR products : BphR2 Part I
08/20/13
- Electrophoresis of PCR products : BphR2 Part I
- Gel purification of PCR products : BphR2 Part I
08/21/13
- Transformation of RBS-BphR2 in DH5α
08/22/13
- PCR of BphR2 Part I
- Electrophoresis of PCR product : BphR2 Part I
- Gel purification of PCR product : BphR2 Part I
08/26/13
- Gibson assembly of RBS-BphR2
- Transformation of RBS-BphR2 in DH5α
08/27/13
- Electrophoresis of RBS-BphR2 Part I, BphR2 Part II
- Electrophoresis of pSB1C3
- Digestion of pSB1C3 by DnpI tto clean it
- Electrophoresis of the digestion of pSB1C3 by DnpI
08/28/13
- Gel purification of pSB1C3 digested by DnpI
- Electrophoresis to check the gel purification of pSB1C3 digested by DnpI
- Gibson assembly of RBS-BphR2
- Transformation of RBS-BphR2 in DH5α
08/29/13
- Colony PCR of RBS-BphR2 in DH5α
08/30/13
- Electrophoresis of Colony PCR of RBS-BphR2
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
08/06/13
- Extraction of BBa_J04450 from DH5α
08/08/13
- Digestion of BBa_J04450 by EcoRI/PstI
- Electrophoresis of BBa_J004450 digested by EcoRI/Pst1 to check if the digestion
- Electrophoresis of BBa_J004450 digested by EcoRI/PstI
- Electroelution of pSB3K3 digested by EcoRI/PstI
08/13/13
- Digestion of BBa_J04450 by EcoRI/PstI
- Electrophoresis to check the digestion of BBa_J04450 by EcoRI/PstI
08/21/13
- Electrophoresis of the digestion of BBa_J04450 by EcoRI/PstI
08/22/13
- Gel purification of the digestion of BBa_J04450 by EcoRI/PstI
- Electrophoresis of gel purification of the digestion of BBa_J04450 by EcoRI/PstI
- Ethanol precipitation of the digestion of BBa_J04450 by EcoRI/PstI
08/23/13
- Electrophoresis to check the ethanol precipitation of the digestion of BBa_J04450 by EcoRI/PstI
08/29/13
- Digestion of BBa_J04450 by EcoRI/PstI
- Electrophoresis to check the digestion of BBa_J04450 by EcoRI/PstI
- Gel purification of the digestion of BBa_J04450 by PstI/SpeI
Characterization of BBa_K1155000
08/07/13
- Transduction of Km in MG1655Z1
08/08/13
- Transduction of Km in MG1655Z1
08/09/13
- Transduction of Km in MG1655Z1 (part II)
08/12/13
- Digestion of BBa_K1155000 by SpeI/PstI, BBa_K1155007 and BBa_K1155003 by XBaI/PstI
- Electrophoresis to check the digestion of BBa_K1155007 and BBa_K1155003 by XBalI/PstI
- Electrophoresis to check the digestion of BBa_K1155000 by SpeI/PstI
- Digestion of BBa_K1155000 by SpeI/PstI
08/13/13
- Electrophoresis to check the digestion of BBa_K1155000 by SpeI/PstI
- Digestion of BBa_K1155000 by SpeI and by SpeI/EcoRI
- Electrophoresis to check the digestion of BBa_K1155000 by SpeI and EcoRI/SpeI
- Liquid culture of MG1655Z1 Δfnr::Km
08/14/13
- Extraction of BBa_K1155000 from DH5α
- Digestion of BBa_K1155000 by SpeI and EsoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155000 by SpeI
- Electrophoresis to check the digestion of BBa_K1155000 by EcoRI/SpeI
- Gel purification of the digestion of BBa_K1155000 by EcoRI/SpeI
- Electrophoresis of the digestion of BBa_K1155003, BBa_K1155007 by XBaI/PstI
- Gel purification of the digestion of BBa_K1155003 and Bba_K1155007 by XBaI/PstI
- Glycerol stock of MG1655Z1 Δfnr::Km
08/19/13
- EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
- Make competent cells of strain MG1655Z1 Δfnr::Km
- Transformation of pcp20 into strain MG1655Z1 Δfnr::Km
08/21/13
- Culture of BBa_K1155000
08/22/13
- Plasmid extraction of BBa_K1155000 from DH5α
- Digestion of BBa_K1155000 by SpeI
08/26/13
- Digestion of BBa_K1155000 by SpeI/PstI
- Denaturation of SpeI/PstI used for the digestion of BBa_K1155000
- Ligation of Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Transformation of ligation of Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Purification colony from strain MG1655Z1 Δfnr::Km containing plasmid pcp20
08/27/13
- Colony PCR of ligation Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Electrophoresis of the colony PCR products : Pndh* with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Streak of Pndh* with RBS-LacZ-Term in pSB1C3 and Pfnr with RBS-Amil CP-Term in pSB1C3 to purify them
- Culture of strain MG1655Z1 Δfnr::Km containing plasmid pcp20
08/28/13
- Purification of colony transformed with ligation : Pndh* with RBS-LacZ-Term in pSB1C3, Pndh* with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions
- Culture of mutant strain MG1655Z1 Δfnr with LB
08/29/13
- Purification colony of strain MG1655Z1 Δfnr with kanamycin, chloramphenicol, ampicilin or LB
- Purification of colony transformed with ligation : Pndh* with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions
- Colony PCR of Pndh* wit RBS_AmilCP-Term in DH5α
- Digestion of BBa_K1155000, BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Electrophoresis to check the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Gel purification of the digestion of Bba_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Electrophoresis to check the gel purification of the digestion of BBa_K1155000 by PstI/SpeI, BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Liquid culture in aerobic or anaerobic conditions of Pndh* with RBS-Amil CP-Term in pSB1C3
WE CHARACTERIZE BBa_K1155000. BBa_K1155003 WORKS TOO.
08/30/13
- Ligation of Pndh* with RBS-Amil CP-Term in pSB1C3
- Transformation of ligation of Pndh* with RBS-Amil CP-Term in pSB1C3 in DH5α
- Ligation of Pndh* with RBS-LacZ-Term in pSB3K3
- Transformation of ligation of Pndh* with RBS-LacZ-Term in pSB3K3 and in DH5α
- Transformation of ligation of Pndh* with RBS-LacZ-Term in pSB3C3 and in DH5α
- Electrophoresis of Colony PCR of Pndh* with RBS_AmilCP-Term in DH5α
- Result of the purification colony of MG1655Z1 Δfnr in ampicilin, chloramphenicol, kanamycin or LB medium [File:Psright.png|20px]]
WE OBTAIN STRAIN MG1655Z1 Δfnr
Characterization BBa_K1155004
08/12/13
- Digestion of BBa_K1155007 and BBa_K1155003 by XBaI/PstI
- Electrophoresis to check the digestion of BBa_K1155007 and BBa_K1155003 by XBalI/PstI
08/13/13
- Extraction of BBa_K1155004 from DH5α
- Digestion of BBa_K1155004 by SpeI and by SpeI/EcoRI
- Electrophoresis to check the digestion of BBa_K1155004 by SpeI
- Electrophoresis to check the digestion of BBa_K1155004 by SpeI/EcoRI
08/14/13
- Gel purification of the digestion of BBa_K1155004 by SpeI
- Extraction of BBa_K1155004 from DH5α
- Digestion of BBa_K1155004 by EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155004 by EcoRI/SpeI
- Digestion of BBa_K1155004 by SpeI and EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155004 by SpeI
- Electrophoresis to check the digestion of BBa_K1155004 by EcoRI/SpeI
- Gel purification of the digestion of BBa_K1155004 by SpeI
- Electrophoresis of the digestion of BBa_K1155003, BBa_K1155007 by XBaI/PstI
- Gel purification of the digestion of BBa_K1155003 and BBa_K1155007 by XBaI/PstI
08/19/13
- EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
- Ligation of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3
- Transformation of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/21/13
- Colony PCR of PnirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/26/13
- Digestion of BBa_K1155004 by SpeI/PstI
- Denaturation of SpeI/PstI used for the digestion of BBa_K1155004
- Ligation of PnirB clone 6 with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Transformation of ligation of PnirB with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
08/27/13
- Colony PCR of ligation PnirB with RBS-LacZ-Term or RBS-Amil CP-Term in pSB1C3
- Electrophoresis of the colony PCR products : PnirB with RBS-LacZ-Term
- Electrophoresis of the colony PCR products : PnirB with RBS-Amil CP-Term in pSB1C3
- Streak of PnirB with RBS-Amil CP-Term in pSB1C3 to purify them
08/28/13
- Purification of colony transformed with ligation : PnirB with RBS-LacZ-Term in pSB1C3, PnirB with RBS-Amil CP-Term in pSB1C3 by streaking in aerobic or anaerobic conditions
08/29/13
- Purification of colony transformed with ligation : PnirB with RBS-LacZ-Term in pSB1C3 by streaking in aerobic or anaerobic conditions
- Electrophoresis to check the digestion of BBa_K1155004 by SpeI/PstI and plasmids already digested by SpeI and after digested by PstI
Characterization of BBa_K1155005
08/12/13
- Digestion of BBa_K1155007 and BBa_K1155003 by XBaI/PstI
- Electrophoresis to check the digestion of BBa_K1155007 and BBa_K1155003 by XBalI/PstI
08/13/13
- Extraction of BBa_K1155005 from DH5α
- Digestion of BBa_K1155005 by SpeI and by SpeI/EcoRI
- Electrophoresis to check the digestion of BBa_K1155005 by SpeI
- Electrophoresis to check the digestion of BBa_K1155005 by SpeI/EcoRI
08/14/13
- Gel purification of the digestion of BBa_K1155005 by SpeI
- Extraction of BBa_K1155005 from DH5α
- Digestion of BBa_K1155005 by EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155005 by EcoRI/SpeI
- Digestion of BBa_K1155005 by SpeI and EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155005 by SpeI
- Electrophoresis to check the digestion of BBa_K1155005 by EcoRI/SpeI
- Gel purification of the digestion of BBa_K1155005 by SpeI
- Electrophoresis of the digestion of BBa_K1155003, BBa_K1155007 by XBaI/PstI
- Gel purification of the digestion of BBa_K1155003 and BBa_K1155007 by XBaI/PstI
08/19/13
- EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
- Ligation of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3
- Transformation of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/21/13
- Colony PCR of PnarG with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/26/13
- Digestion of BBa_K1155005 by SpeI/PstI
- Denaturation of SpeI/PstI used for the digestion of BBa_K1155005
08/29/13
- Electrophoresis to check the digestion of BBa_K1155005 by SpeI/PstI and plasmids already digested by SpeI and after digested by PstI
Characterization of BBa_K1155006
08/12/13
- Digestion of BBa_K1155007 and BBa_K1155003 by XBaI/PstI
- Electrophoresis to check the digestion of BBa_K1155007 and BBa_K1155003 by XBalI/PstI
08/13/13
- Extraction of BBa_K1155006 from DH5α
- Digestion of BBa_K1155006 by SpeI and by SpeI/EcoRI
- Electrophoresis to check the digestion of BBa_K1155006 by SpeI
- Electrophoresis to check the digestion of BBa_K1155006 by SpeI/EcoRI
08/14/13
- Gel purification of the digestion of BBa_K1155006 by SpeI
- Extraction of BBa_K1155006 from DH5α
- Digestion of BBa_K1155006 by EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155006 by EcoRI/SpeI
- Digestion of BBa_K1155006 by SpeI and EcoRI/SpeI
- Electrophoresis to check the digestion of BBa_K1155006 by SpeI and EcoRI/SpeI
- Gel purification of the digestion of BBa_K1155006 by EcoRI/SpeI and SpeI
- Electrophoresis of the digestion of BBa_K1155003, BBa_K1155007 by XBaI/PstI
- Gel purification of the digestion of BBa_K1155003 and BBa_K1155007 by XBaI/PstI
08/19/13
- EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
- Ligation of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3
- Transformation of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/21/13
- Colony PCR of PnarK with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α
08/26/13
- Digestion of BBa_K1155006 by SpeI/PstI
- Denaturation of SpeI/PstI used for the digestion of BBa_K1155006
08/29/13
- Electrophoresis to check the digestion of BBa_K1155006 by SpeI/PstI and plasmids already digested by SpeI and after digested by PstI
- Digestion of BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Electrophoresis to check the digestion of BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Gel purification of the digestion of BBa_K1155003, BBa_K1155007 by Xbal/PstI
- Electrophoresis to check the gel purification of the digestion of BBa_K1155003, BBa_K1155007 by Xbal/PstI
08/30/13
- Ligation of PnarK with RBS-LacZ-Term in pSB3K3
- Transformation of ligation of PnarK with RBS-LacZ-Term in pSB3K3 in DH5α
BBa_K1155006 WORKS IN ANAEROBIC CONDITIONS. BBa_K1155007 WORKS.
Written by Nadia
