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Team:Warsaw/Calendar-Main/11 July 2009
From 2009.igem.org
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| - | ====Gel out phoP/phoQ | + | ====Gel out phoP/phoQ [Kama]==== |
*Isolation of fragment of the correct lenght(˜2200bp)from the gel was performed with the [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf A&A "Gel-out" kit]. | *Isolation of fragment of the correct lenght(˜2200bp)from the gel was performed with the [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf A&A "Gel-out" kit]. | ||
| - | ==== Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 | + | ==== Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2 [Ania] ==== |
Tasks: | Tasks: | ||
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! DNA sample | ! DNA sample | ||
! restriction enzymes | ! restriction enzymes | ||
| - | ! expected fragments | + | ! expected fragments [bp] |
|- | |- | ||
| R0051(pcI) on pSB1A2 | | R0051(pcI) on pSB1A2 | ||
| PvuI HindII | | PvuI HindII | ||
| - | | | + | | 695, 1447 |
|} | |} | ||
| - | ====Strain testing constructs - Franek==== | + | ====Strain testing constructs - [Franek]==== |
* Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks: | * Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks: | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 BBa_R0051] - AraC inhibited promoter | [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 BBa_R0051] - AraC inhibited promoter | ||
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! DNA sample | ! DNA sample | ||
! restriction enzymes | ! restriction enzymes | ||
| - | ! expected fragments | + | ! expected fragments [bp] |
|- | |- | ||
| LacI on pSB1A2 | | LacI on pSB1A2 | ||
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| - | ====Monika ==== | + | ====Franek lub Monika opisze minilizy dla Moniki ==== |
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{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Revision as of 23:04, 11 July 2009
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Contents |
Gel out phoP/phoQ [Kama]
- Isolation of fragment of the correct lenght(˜2200bp)from the gel was performed with the [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf A&A "Gel-out" kit].
Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2 [Ania]
Tasks:
- Transformation of chemocompetent strain of E. with [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid taken out of the distribution 200 Kit Plate 1 well 2O.
- Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks:
[http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 BBa_R0051] - promoter (lambda cI regulated); [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032] - RBS.3 (medium)
- Digestion to confirm plasmid extraction. DNA that should contain R0051 on pSB1A2 plasmid was digested with PvuI and HindII. Digestion mix contained 5ul of extracted DNA, 2ul of Fermentas BamHI buffer , 0.3ul of each enzyme and water added to obtain 20ul total volume.
| DNA sample | restriction enzymes | expected fragments [bp] |
|---|---|---|
| R0051(pcI) on pSB1A2 | PvuI HindII | 695, 1447 |
Strain testing constructs - [Franek]
- Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks:
[http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 BBa_R0051] - AraC inhibited promoter [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032] - LacI inhibited promoter
- Digestion to confirm plasmid extraction. DNA that should contain pAraC on pSB1A2 plasmid was digested with BamHI PvuI, pLacI on pSB1A2 was digested with IPvuI and PvuII . Digestion mix contained 5ul of extracted DNA, 2ul of Fermentas BamHI buffer , 0.3ul of each enzyme and water added to obtain 20ul total volume.
| DNA sample | restriction enzymes | expected fragments [bp] |
|---|---|---|
| LacI on pSB1A2 | PvuI PvuII | 728, 1351 |
| pArac on pSB1A2 | BamHI PvuI | 782, 1446 |
Franek lub Monika opisze minilizy dla Moniki
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