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From 2011.igem.org

Contents 1 Cyrille 2 Danyel 2.1 Miniprep & Glycerols 2.2 Digestion 2.3 GFP amber 2.4 Transformation 2.5 To Do List 3 Ouriel, Hovannes, Baptiste 3.1 Preparation of slides 3.2 Observation

Cyrille

Preparation of the nexts cloning steps. Making the miniprep for the S69, S102, S103, S104, S105.

Making the glycerols for S102, S103, S104, S105

Transforming S101 and S102 for minipreping and glycerol. Plaated and growth overnight.

Danyel

Miniprep & Glycerols

• S69 (pHM3)
• S92 (Pveg_spoVG_tRNA_TT in pHM3) [Sequencing]
• S98 1,2,3,6 (pHS_tRNA) [Sequencing]
• S99 1,2,3,4 (pHS_tRNA_TT) [Sequencing]

Digestion

Everything was digested on E & P

• S69 - all 5 minipreps stored in the fridge were digested. (There seems to be a problem with the 3 pHM3 minipreps that were made today)
• S79 (pT7_RBS_GFP_TT)
• S98
• S99
• S27 (pSB1C3)
• S105 (Pveg_spoVG_T7amber_TT)

All bands of the appropriate sizes were extracted and purified.

GFP amber

Quickchange mutagenesis PCR has been run.

Transformation

Transformation of S38 in S24. Along with the 5 ligations tubes of Babak.

To Do List

• Check transformation results. Eventually follow-up with a PCR colony, then sequencing.
• Digest GFP quickchange products with DpnI enzyme.
• Check sequencing results for S92 S98 S99
• Read with Tecan concentration of all gel extractions.
• Ligate & Transform : S79 into S27 and S79 into pHM3, S98 into S69, S99 into S69, S105 into S27

Ouriel, Hovannes, Baptiste

Preparation of slides

Mix of 3610 & 3610-GFP on one well. Negative control on the second well. Both cultures were at 0,8 OD when we centrifuged and concentrated them in 150 micro liters of LB.

Observation

We could not interpret the pictures we took because of the too short time lapse we observed the cells. Plus, some of our observation were at stationnary phase which probably could avoid cells to interact.