Team:Paris Liliane Bettencourt/Notebook/2011/07/28/

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Team IGEM Paris 2011

Contents

Cyrille

The overnight pHM3 QCM PRC was collected, and digested. The results where loaded on a gel, before and after the digestion. The results are shown below:

Ladder ; Ctrl - bef. digest ; tube 1 bef. digest ; tube 2 bef. digest ; crtl - after digest ; tube 1 after digest ; tube 2 after digest

The results are positive. The supercoiled template DNA appears upper thant the PCR band. After the digestion, the upper band smoothen and become weaker, and even disapears in the tube two. A bit longer digestion was proposed to Danyel for his own QCM. It seems that the PCR have worked so the transformation was carried on.

It is surprising that we dont see the ctrl - band. We didn't went farther on this problem, because the template band was clearly visible and identifiable.

Camille & Danyel

T7 amber

• Quick Mutation PCR product was digested directly within the PCR tube with 1.5µL of DpnI (FastDigest) for 20' at 37ºC • The digestion was followed by a PCR purification.

Final concentration : 419.7 ng/µL with a ratio: 1.94

• Transformation: 20µL and 200µL for each of the following: - control + : B7 - T7 amber in LBA + Cm plates - (Axel) Cx in LBA + Amp plates incubation overnight at 37ºC (start 19:55)

tRNA amber suppressor

• Transformation of the KtRNA strains was successful.
• Miniprep of 6 clones were prepared.

Kevin

• Transformation: E. Coli with 2 plasmids
pSG20 - pDAG464
pFX234 - pDAG479
Control Amp

Results : Didn't grow ! Maybe competents cells are not efficient.

• Cultur overnight from glycerol : pSG20 - pDAG464 - pFX234 - pDAG479

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