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Team:Bonn/Notebook
From 2012.igem.org
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=== May 2012 === | === May 2012 === | ||
| - | ==== | + | ==== 23.05 ==== |
SOC medium and SOB medium prepared, microbial culture: DH5α | SOC medium and SOB medium prepared, microbial culture: DH5α | ||
Agar plates, chemo-competent DH5α | Agar plates, chemo-competent DH5α | ||
| - | ==== | + | ==== 31.05 ==== |
First successful transformation of iGEM plasmids. XL1-blue bacteria was much more competent than our DH5α, so we used these for now. | First successful transformation of iGEM plasmids. XL1-blue bacteria was much more competent than our DH5α, so we used these for now. | ||
| - | ==== | + | |
| + | === June 2012 === | ||
| + | |||
| + | ==== 04.06 ==== | ||
First Midi-Preps of iGEM plasmids from our distribution kit. | First Midi-Preps of iGEM plasmids from our distribution kit. | ||
==== 06.06 - 06.08 ==== | ==== 06.06 - 06.08 ==== | ||
| - | + | First restrictions, some of them were successful. The failure on our LOVTap plasmid was misinterpreted: We thought we had problems with the restriction, PCR or our transformation, but in fact the iGEM LOV biobrick was broken. | |
| - | === | + | ==== 08.06 - 03.07 ==== |
| - | + | PCR, transformation and restriction troubleshooting to find the issue in our LOVTap transformation. All tests and modifications of our protocols failed for the LOVTap plasmid, but worked in with other biobricks and sample plasmids. So we finally send the LOVTap iGEM plasmid to a company for sequencing. | |
| - | + | ||
=== July 2012 === | === July 2012 === | ||
| - | : | + | ==== 04.07 ==== |
| + | Sequencing results of all our biobricks are available: Sequence is okay for RBS32, LacZα, pLac and our double-terminator. Frameshift found in lacI plasmid. | ||
| + | LOVTap was garbage. | ||
| + | |||
| + | ==== later this moths... ==== | ||
| + | Further troubleshooting and optimization of our standard procedures. We also tried to figure out if only our distribution plate contained the broken LOV, so we asked other iGEM teams to provide us with samples from their plates. We did transformations and sequencing, but these LOV plasmids showed the same pattern of degeneration our LOV did. | ||
=== August 2012 === | === August 2012 === | ||
| - | + | ==== 06.08 ==== | |
| + | First application of the 3A assembly, following the official iGEM protocol. (we used a different one before). | ||
| + | * Restrictions | ||
| + | * Ligations | ||
| + | |||
| + | ==== 07.08 ==== | ||
| + | Transformation of constructs prepared the day before. | ||
| + | |||
| + | ==== 08.08 ==== | ||
| + | Restriction analysis of constructs LacZα-TT-pSB1C3 and ccdB-TT-pSB1C3 | ||
| + | ==== 10.08 ==== | ||
| + | Sequencing of created LacZα-TT-pSB1C3 and ccdB-TT-pSB1C3 constructs for verification. | ||
=== September 2012 === | === September 2012 === | ||
::TODO | ::TODO | ||
Revision as of 20:47, 15 September 2012
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Contents |
Lab Notebook
Everything we did in our lab, including the standard protocols we used.
May 2012
23.05
SOC medium and SOB medium prepared, microbial culture: DH5α Agar plates, chemo-competent DH5α
31.05
First successful transformation of iGEM plasmids. XL1-blue bacteria was much more competent than our DH5α, so we used these for now.
June 2012
04.06
First Midi-Preps of iGEM plasmids from our distribution kit.
06.06 - 06.08
First restrictions, some of them were successful. The failure on our LOVTap plasmid was misinterpreted: We thought we had problems with the restriction, PCR or our transformation, but in fact the iGEM LOV biobrick was broken.
08.06 - 03.07
PCR, transformation and restriction troubleshooting to find the issue in our LOVTap transformation. All tests and modifications of our protocols failed for the LOVTap plasmid, but worked in with other biobricks and sample plasmids. So we finally send the LOVTap iGEM plasmid to a company for sequencing.
July 2012
04.07
Sequencing results of all our biobricks are available: Sequence is okay for RBS32, LacZα, pLac and our double-terminator. Frameshift found in lacI plasmid. LOVTap was garbage.
later this moths...
Further troubleshooting and optimization of our standard procedures. We also tried to figure out if only our distribution plate contained the broken LOV, so we asked other iGEM teams to provide us with samples from their plates. We did transformations and sequencing, but these LOV plasmids showed the same pattern of degeneration our LOV did.
August 2012
06.08
First application of the 3A assembly, following the official iGEM protocol. (we used a different one before).
* Restrictions * Ligations
07.08
Transformation of constructs prepared the day before.
08.08
Restriction analysis of constructs LacZα-TT-pSB1C3 and ccdB-TT-pSB1C3
10.08
Sequencing of created LacZα-TT-pSB1C3 and ccdB-TT-pSB1C3 constructs for verification.
September 2012
- TODO
