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Contents 1 Lab Notebook 1.1 Summary 1.2 Highlights by date 1.2.1 May 2012 1.2.1.1 23.05 1.2.1.2 31.05 1.2.2 June 2012 1.2.2.1 04.06 1.2.2.2 06.06 - 06.08 1.2.2.3 08.06 - 03.07 1.2.3 July 2012 1.2.3.1 04.07 1.2.3.2 later this month... 1.2.4 August 2012 1.2.4.1 06.08 1.2.4.2 07.08 1.2.4.3 08.08 1.2.4.4 10.08 1.2.5 September 2012 1.3 Protocols

Lab Notebook

Everything we did in our lab, including the standard protocols we used.

Summary

TODO

Highlights by date

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 month...

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

Protocols

TODO

(Protocols will have to be linked here, please don't add the big texts directly!)