"
Team:Warsaw/Calendar-Main/11 July 2009
From 2009.igem.org
(Difference between revisions)
| Line 2: | Line 2: | ||
<!-- do not edit above me! --> | <!-- do not edit above me! --> | ||
| - | ====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: | ||
| Line 20: | Line 20: | ||
! 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 | ||
| Line 39: | Line 39: | ||
! DNA sample | ! DNA sample | ||
! restriction enzymes | ! restriction enzymes | ||
| - | ! expected fragments | + | ! expected fragments [bp] |
|- | |- | ||
| LacI on pSB1A2 | | LacI on pSB1A2 | ||
| Line 51: | Line 51: | ||
| - | ====Monika ==== | + | ====Franek lub Monika opisze minilizy dla Moniki ==== |
<!-- do not remove this! --> | <!-- do not remove this! --> | ||
{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Revision as of 23:04, 11 July 2009
 |  |
 |
  |
 |
|
Contents |
Gel out phoP/phoQ [Kama]
- Isolation of fragment of the correct lenght(˜2200bp)from the gel was performed with the A&A "Gel-out" kit.
Construction of K177012 operon1_part2 [Ania]
Tasks:
- Transformation of chemocompetent strain of E. with 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:
BBa_R0051 - promoter (lambda cI regulated); 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:
BBa_R0051 - AraC inhibited promoter 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
|
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
