Week 9
From 2007.igem.org
(Difference between revisions)
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::'''08/28/07''' | ::'''08/28/07''' | ||
*Lab meeting on: | *Lab meeting on: | ||
- | -model | + | -model; |
- | -LacI cloning under the pTetR [http://partsregistry.org/Part:BBa_R0040 R0040])promoter control | + | -LacI cloning under the pTetR [http://partsregistry.org/Part:BBa_R0040 R0040])promoter control. |
*We transform new biobricks:[http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] | *We transform new biobricks:[http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] | ||
- | * We inoculate | + | * We inoculate [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] colonies in 5ml; |
* There are no colonies for [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412]([http://partsregistry.org/Part:BBa_I763010 (I763010)] ligation. | * There are no colonies for [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412]([http://partsregistry.org/Part:BBa_I763010 (I763010)] ligation. | ||
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*[http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] digestions with Xba/Pst1. | *[http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] digestions with Xba/Pst1. | ||
*[http://partsregistry.org/Part:BBa_B0034 B0034] digestion with Spe/Pst1. | *[http://partsregistry.org/Part:BBa_B0034 B0034] digestion with Spe/Pst1. | ||
- | *Run on | + | *Run on electrophoresis gel and [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034] band extraction. We have some problems with [http://partsregistry.org/Part:BBa_J06550 J06550] plasmid vector due to its molecular weight (too heavy) and with [http://partsregistry.org/Part:BBa_S0103 S0103] insert for the same reason (too heavy). Thus, we decide to use another biobrick. |
*Ligation for [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] ([http://partsregistry.org/Part:BBa_I763016 I763016]), [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sendbox),[http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] ([http://partsregistry.org/Part:BBa_I763020 I763020]). | *Ligation for [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] ([http://partsregistry.org/Part:BBa_I763016 I763016]), [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sendbox),[http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] ([http://partsregistry.org/Part:BBa_I763020 I763020]). | ||
*We transform the ligations. | *We transform the ligations. |
Revision as of 10:36, 5 September 2007
- 08/27/07
- New R0051+P0412 (I763010) ligation with a new protocol (3h 15°C and 1.2ul Ligase).
- We transform R0051+P0412 (I763010) ligation and new S03520, S0100, B0034, J06550, S0103 biobricks.
- 08/28/07
- Lab meeting on:
-model;
-LacI cloning under the pTetR R0040)promoter control.
- 08/29/07
- Miniprep of S03520, S0100, B0034, J06550, S0103.
- S03520, S0100, J06550, S0103 digestions with Xba/Pst1.
- B0034 digestion with Spe/Pst1.
- Run on electrophoresis gel and S03520, S0100, B0034 band extraction. We have some problems with J06550 plasmid vector due to its molecular weight (too heavy) and with S0103 insert for the same reason (too heavy). Thus, we decide to use another biobrick.
- Ligation for I763012 + J06550 (I763016), I763012 + S0103 (missed in the sendbox),B0034 + J22101 (missed in the sendbox), B0034 + J04631 (I763020).
- We transform the ligations.
- 08/30/07
- There are colonies in the 07/30/07 plates.
- We inoculate a E0020 and R0040 colony in 5ml.
- We perform some tests to set up the fluorescence acquisition with the PMT (photomultiplier tube).
- 08/31/07
J22101 (missed in the sandbox), B0034 + J04631 (I763020), E0020 and of R0040.
B0034 + J22101 digestion with Xba/Pst1; B0034 + J04631 (I763020) digestion with Xba/Pst1;