Week 9
From 2007.igem.org
(Difference between revisions)
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*[http://partsregistry.org/Part:BBa_B0034 B0034] digestion with Spe/Pst1. | *[http://partsregistry.org/Part:BBa_B0034 B0034] digestion with Spe/Pst1. | ||
*Run on electroforesi 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 notice problems with [http://partsregistry.org/Part:BBa_J06550 J06550] band height (the insert is too heavy) and with [http://partsregistry.org/Part:BBa_S0103 S0103] band because the plasmid too heavy, however we extract insert bands for both. | *Run on electroforesi 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 notice problems with [http://partsregistry.org/Part:BBa_J06550 J06550] band height (the insert is too heavy) and with [http://partsregistry.org/Part:BBa_S0103 S0103] band because the plasmid too heavy, however we extract insert bands for both. | ||
- | *Ligation with a new protocol of I763012 + [http://partsregistry.org/Part:BBa_J06550 J06550] (I763016), I763012 + S0103 (missed in the sendbox),[http://partsregistry.org/Part:BBa_B0034 B0034] + J22101 (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + J04631 (I763020). | + | *Ligation with a new protocol of [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 10:58, 31 August 2007
- 08/27/07
- New R0051+P0412 (I763010) ligation with a new protocol (3h 15°C and 1.2 ul Ligasi).
- Transformation of R0051+P0412 (I763010) ligation and of new biobricks S03520, S0100, B0034, J06550, S0103.
- 08/28/07
- We hold a meeting and the topics of the day are:
-model agreed;
-we agree the position of LacI not controlled by Plac but by costitutionaly expressed as PTetR gene (R0040);
-reseach of genomic pLacI sequence to draw the primers or to order the synthesis.
- We transform new biobricks:E0020 (to put after CMV J52034 promoter) and R0040 (whith witch adjust LacI expression);
- 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 electroforesi gel and S03520, S0100, B0034 band extraction. We notice problems with J06550 band height (the insert is too heavy) and with S0103 band because the plasmid too heavy, however we extract insert bands for both.
- Ligation with a new protocol of 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 all ligations.
- We inoculate a ligations, E0020 and R0040 colony in 5 ml.
- Dimension tests with PMT to set gain and dimension area.
- 08/31/07
J22101 (missed in the sendbox), B0034 + J04631 (I763020), E0020 and of R0040.